Receptor that binds IL-17

ABSTRACT

Isolated receptors for IL-17, DNA&#39;s encoding such receptors, and pharmaceutical compositions made therefrom, are disclosed. The isolated receptors can be used to regulate an immune response.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a divisional application of U.S. patent applicationSer. No. 08/620,694, filed Mar. 21, 1996, now allowed as U.S. Pat. No.5,869,286, which is a continuation-in-part of U.S. application Ser. No.08/538,765, filed Aug. 7, 1995, now abandoned, which is acontinuation-in-part of U.S. application Ser. No. 08/410,535, filed Mar.23, 1995.

TECHNICAL FIELD OF THE INVENTION

The present invention relates generally to the field of cytokinereceptors, and more specifically to cytokine receptor proteins havingimmunoregulatory activity.

BACKGROUND OF THE INVENTION

Cytokines are hormone-like molecules that regulate various aspects of animmune or inflammatory response. Cytokines exert their effects byspecifically binding receptors present on cells, and transducing asignal to the cells. Rouvier et al. (J. Immunol. 150:5445; 1993)reported a novel cDNA which they termed CTLA-8. The putative CTLA8protein is 57% homologous to the predicted amino acid sequence of anopen reading frame (ORF) present in Herpesvirus saimiri (HSV) referredto as HVS13 (Nicholas et al. Virol. 179:1 89, 1990; Albrecht et al., J.Virol. 66:5047;1992). However, the function, if any of either CTLA-8 orHVS13 was not known, nor was a receptor or binding protein for CTLA-8 orHVS13 known. Thus, prior to the present invention, there was a need inthe art to determine the function of CTLA-8 and HVS13, and to identifyreceptor molecules or binding proteins that play a role in the functionof these proteins.

SUMMARY OF THE INVENTION

The present invention identifies a novel receptor that binds IL-17(CTLA-8) and HVS13, a viral homolog of IL-17; DNAs encoding the novelreceptor and novel receptor proteins are provided. The receptor is aType I transmembrane protein; the mouse receptor has 864 amino acidresidues, the human receptor has 866 amino acid residues. Soluble formsof the receptor can be prepared and used to regulate immune responses ina therapeutic setting; accordingly, pharmaceutical compositionscomprising soluble forms of the novel receptor are also provided.Deleted forms and fusion proteins comprising the novel receptor, andhomologs thereof are also disclosed. Also provided are methods ofregulating an immune response, and methods of suppressing rejection ofgrafted organs or tissue. These and other aspects of the presentinvention will become evident upon reference to the following detaileddescription of the invention.

DETAILED DESCRIPTION OF THE INVENTION

A soluble IL-17 (CTLA-8) protein and an ORF present in Herpesvirussaimiri (HVS13) were expressed as fusion proteins comprising animmunoglobulin Fc region, and used to screen cells for expression of areceptor for IL-17. T cell thymoma EL4 cells were found to bind theHVS13/Fc as well as murine CTLA8 (IL-17)/Fc fusion protein. A cDNAlibrary from EL4 cells was prepared and screened for expression of thereceptor. The receptor is a Type I transmembrane protein with 864 aminoacid residues, which is referred to as IL-17R (CTLA-8R). Various formsof IL-17R were prepared, including IL-17R/Fc protein, a soluble IL-17Rwhich contains the signal peptide and extracellular domain of IL-17R,and a soluble IL-17R/Flag® construct. A human IL-17R was isolated from ahuman peripheral blood lymphocyte library by cross-specieshybridization, and exhibits similarities to the murine IL-17R.

IL-17. HVS13 and Homologous Proteins

CTLA-8 refers to a cDNA cloned from an activated T cell hybridoma clone(Rouvier et al., J. Immunol. 150:5445; 1993). Northern blot analysisindicated that CTLA-8 transcription was very tissue specific. The CTLA-8gene was found to map at chromosomal site 1a in mice, and at 2q31 inhumans. Although a protein encoded by the CTLA-8 gene was neveridentified by Rouvier et al, the predicted amino acid sequence of CTLA-8was found to be 57% homologous to the predicted amino acid sequence ofan ORF present in Herpesvirus Sairniri, HVS13. The CTLA-8 protein isreferred to herein as Interleukin-17 (IL-17).

The complete nucleotide sequence of the genome of HVS has been reported(Albrecht et al., J. Virol. 66:5047; 1992). Additional studies on one ofthe HVS open reading frames (ORFs), HVS13, are described in Nicholas etal., Virol. 179:1 89; 1990. HVS13 is a late gene which is present in theHind III-G fragment of HVS. Antisera developed against peptides derivedfrom HVS13 are believed to react with a late protein (Nicholas et al.,supra).

As described U.S. Ser. No. 08/462,353, a CIP of U.S. Ser. No.08/410,536, filed Mar. 23, 1995, full length murine CTLA-8 protein and aCTLA-8/Fc fusion protein were expressed, tested, and found to act as acostimulus for the proliferation of T cells. Human IL-17 (CTLA-8) wasidentified by probing a human T cell library using a DNA fragmentderived from degenerate PCR; homologs of IL-17 (CTLA-8) are expected toexist in other species as well. A full length HVS13 protein, as well asan HVS13/Fc fusion protein, were also expressed, and found to act in asimilar manner to IL-17 (CTLA-8) protein. Moreover, other species ofherpesviruses are also likely to encode proteins homologous to thatencoded by HVS13.

Proteins and Analogs

The present invention provides isolated IL-17R and homologs thereofhaving immunoregulatory activity. Such proteins are substantially freeof contaminating endogenous materials and, optionally, withoutassociated native-pattern glycosylation. Derivatives of IL-17R withinthe scope of the invention also include various structural forms of theprimary protein which retain biological activity. Due to the presence ofionizable amino and carboxyl groups, for example, an IL-17R protein maybe in the form of acidic or basic salts, or may be in neutral form.Individual amino acid residues may also be modified by oxidation orreduction.

The primary amino acid structure may be modified by forming covalent oraggregative conjugates with other chemical moieties, such as glycosylgroups, lipids, phosphate, acetyl groups and the like, or by creatingamino acid sequence mutants. Covalent derivatives are prepared bylinking particular functional groups to amino acid side chains or at theN- or C-ternuni.

Soluble forms of IL-17R are also within the scope of the invention. Thenucleotide and predicted amino acid sequence of the murine IL-17R isshown in SEQ ID NOs:1 and 2. Computer analysis indicated that theprotein has an N-terminal signal peptide with a cleavage site betweenamino acid 31 and 32. Those skilled in the art will recognize that theactual cleavage site may be different than that predicted by computeranalysis. Thus, the N-terminal amino acid of the cleaved peptide isexpected to be within about five amino acids on either side of thepredicted cleavage site. The signal peptide is followed by a 291 aminoacid extracellular domain, a 21 amino acid transmembrane domain, and a521 amino acid cytoplasmic tail. Soluble IL-17R comprises the signalpeptide and the extracellular domain (residues 1 to 322 of SEQ ID NO:1)or a fragment thereof. Alternatively, a different signal peptide can besubstituted for residues 1 through 3 1 of SEQ ID NO:1.

The nucleotide and predicted amino acid sequence of the human IL-I17R isshown in SEQ ID NOs:9 and 10. It shares many features with the murineIL-17 R. Computer analysis indicated that the protein has an N-terminalsignal peptide with a cleavage site between amino acid 27 and 28. Thoseskilled in the art will recognize that the actual cleavage site may bedifferent than that predicted by computer analysis. Thus, the N-terminalamino acid of the cleaved peptide is expected to be within about fiveamino acids on either side of the predicted cleavage site. The signalpeptide is followed by a 293 amino acid extracellular domain, a 21 aminoacid transmembrane domain, and a 525 amino acid cytoplasmic tail.Soluble IL-17R comprises the signal peptide and the extracellular domain(residues 1 to 320 of SEQ ID NO:1) or a fragment thereof. Alternatively,a different signal peptide can be substituted for the native signalpeptide.

Other derivatives of the IL-17R protein and homologs thereof within thescope of this invention include covalent or aggregative conjugates ofthe protein or its fragments with other proteins or polypeptides, suchas by synthesis in recombinant culture as N-terminal or C-terminalfusions. For example, the conjugated peptide may be a signal (or leader)polypeptide sequence at the N-terminal region of the protein whichco-translationally or post-translationally directs transfer of theprotein from its site of synthesis to its site of function inside oroutside of the cell membrane or wall (e.g., the yeast α-factor leader).

Protein fusions can comprise peptides added to facilitate purificationor identification of IL-17R proteins and homologs (e.g., poly-His). Theamino acid sequence of the inventive proteins can also be linked to anidentification peptide such as that described by Hopp et al.,Bio/Technology 6:1204 (1988). Such a highly antigenic peptide providesan epitope reversibly bound by a specific monoclonal antibody, enablingrapid assay and facile purification of expressed recombinant protein.The sequence of Hopp et al. is also specifically cleaved by bovinemucosal enterokinase, allowing removal of the peptide from the purifiedprotein. Fusion proteins capped with such peptides may also be resistantto intracellular degradation in E. coli.

Fusion proteins further comprise the amino acid sequence of a IL-17Rlinked to an immunoglobulin Fc region. An exemplary Fc region is a humanIgG1 having a nucleotide and amino acid sequence set forth in SEQ IDNO:4. Fragments of an Fc region may also be used, as can Fc muteins suchas those described in U.S. Ser. No. 08/145,830, filed Oct. 29, 1993.Depending on the portion of the Fc region used, a fusion protein may beexpressed as a dimer, through formation of interchain disulfide bonds.If the fusion proteins are made with both heavy and light chains of anantibody, it is possible to form a protein oligomer with as many as fourIL-17R regions.

In another embodiment, IL-17R and homologs thereof further comprise anoligomerizing zipper domain. Zipper domains are described in U.S. Ser.No. 08/107,353, filed Aug. 13, 1993, the relevant disclosure of which isincorporated by reference herein. Examples of leucine zipper domains arethose found in the yeast transcription factor GCN4 and a heat-stableDNA-binding protein found in rat liver (C/EBP; Landschulz et al.,Science 243:1681, 1989), the nuclear transforming proteins, fos and jun,which preferentially form a heterodimer (O'Shea et al., Science 245:646,1989; Turner and Tjian, Science 243:1689, 1989), and the gene product ofthe murine proto-oncogene, c-myc (Landschulz et al., Science 240:1759,1988). The fusogenic proteins of several different viruses, includingparamyxovirus, coronavirus, measles virus and many retroviruses, alsopossess leucine zipper domains (Buckland and Wild, Nature 338:547, 1989;Britton, Nature 353:394, 1991; Delwart and Mosialos, AIDS Research andHuman Retroviruses 6:703, 1990).

Derivatives of IL-17R may also be used as immunogens, reagents in invitro assays, or as binding agents for affinity purification procedures.Such derivatives may also be obtained by cross-linking agents, such asM-maleimidobenzoyl succinimide ester and N-hydroxysuccinimnide, atcysteine and lysine residues. The inventive proteins may also becovalently bound through reactive side groups to various insolublesubstrates, such as cyanogen bromide-activated, bisoxirane-activated,carbonyldiimidazole-activated or tosyl-activated agarose structures, orby adsorbing to polyolefin surfaces (with or without glutaraldehydecross-linking). Once bound to a substrate, proteins may be used toselectively bind (for purposes of assay or purification) antibodiesraised against the IL-17R or against other proteins which are similar tothe IL-17R, as well as other proteins that bind IL-17R or its homologousproteins.

The present invention also includes IL-17R with or without associatednative-pattern glycosylation. Proteins expressed in yeast or mammalianexpression systems, e.g., COS-7 cells, may be similar or slightlydifferent in molecular weight and glycosylation pattern than the nativemolecules, depending upon the expression system. Expression of DNAsencoding the inventive proteins in bacteria such as E. coli providesnon-glycosylated molecules. Functional mutant analogs of IL-17R proteinor homologs thereof having inactivated N-glycosylation sites can beproduced by oligonucleotide synthesis and ligation or by site-specificmutagenesis techniques. These analog proteins can be produced in ahomogeneous, reduced-carbohydrate form in good yield using yeastexpression systems. N-glycosylation sites in eukaryotic proteins arecharacterized by the an-ino acid triplet Asn-A₁ -Z, where A₁ is anyamino acid except Pro, and Z is Ser or Thr. In this sequence, asparagineprovides a side chain amino group for covalent attachment ofcarbohydrate. Such a site can be eliminated by substituting anotheramino acid for Asn or for residue Z, deleting Asn or Z, or inserting anon-Z amino acid between A₁ and Z, or an amino acid other than Asnbetween Asn and A₁.

IL-17R protein derivatives may also be obtained by mutations of thenative IL-17R or its subunits. A IL-17R mutated protein, as referred toherein, is a polypeptide homologous to a IL-17R protein but which has anamino acid sequence different from the native IL-17R because of one or aplurality of deletions, insertions or substitutions. The effect of anymutation made in a DNA encoding a IL-17R peptide may be easilydetermined by analyzing the ability of the mutated IL-17R peptide toinhibit costimulation of T or B cells by IL-17 (CTLA-8) or homologousproteins, or to bind proteins that specifically bind IL-17R (forexample, antibodies or proteins encoded by the CTLA-8 cDNA or the HVS13ORF). Moreover, activity of IL-17R analogs, muteins or derivatives canbe determined by any of the assays methods described herein. Similarmutations may be made in homologs of IL-17R, and tested in a similarmanner.

Bioequivalent analogs of the inventive proteins may be constructed by,for example, making various substitutions of residues or sequences ordeleting terminal or internal residues or sequences not needed forbiological activity. For example, cysteine residues can be deleted orreplaced with other amino acids to prevent formation of incorrectintramolecular disulfide bridges upon renaturation. Other approaches tomutagenesis involve modification of adjacent dibasic amino acid residuesto enhance expression in yeast systems in which KEX2 protease activityis present.

Generally, substitutions should be made conservatively; i.e., the mostpreferred substitute amino acids are those which do not affect theability of the inventive proteins to bind their ligands in a mannersubstantially equivalent to that of native mIL-17R or hIL-17R. Examplesof conservative substitutions include substitution of amino acidsoutside of the binding domain(s), and substitution of amino acids thatdo not alter the secondary and/or tertiary structure of IL-17R andhomologs thereof. Additional examples include substituting one aliphaticresidue for another, such as Ile, Val, Leu, or Ala for one another, orsubstitutions of one polar residue for another, such as between Lys andArg; Glu and Asp; or Gln and Asn. Other such conservative substitutions,for example, substitutions of entire regions having similarhydrophobicity characteristics, are well known.

Similarly, when a deletion or insertion strategy is adopted, thepotential effect of the deletion or insertion on biological activityshould be considered. Subunits of the inventive proteins may beconstructed by deleting terminal or internal residues or sequences.Fragments of IL-17R that bind IL-17 can be readily prepared (forexample, by using restriction enzymes to delete portions of the DNA) andtested for their ability to bind IL-17. Additional guidance as to thetypes of mutations that can be made is provided by a comparison of thesequence of IL-17R to proteins that have similar structures, as well asby performing structural analysis of the inventive proteins.

Mutations in nucleotide sequences constructed for expression of analogIL-17R CTLA-8R) must, of course, preserve the reading frame phase of thecoding sequences and preferably will not create complementary regionsthat could hybridize to produce secondary mRNA structures such as loopsor hairpins which would adversely affect translation of the receptormRNA. Although a mutation site may be predetermined, it is not necessarythat the nature of the mutation per se be predetermined. For example, inorder to select for optimum characteristics of mutants at a given site,random mutagenesis may be conducted at the target codon and theexpressed mutated viral proteins screened for the desired activity.

Not all mutations in the nucleotide sequence which encodes a IL-17Rprotein or homolog thereof will be expressed in the final product, forexample, nucleotide substitutions may be made to enhance expression,primarily to avoid secondary structure loops in the transcribed mRNA(see EPA 75,444A, incorporated herein by reference), or to providecodons that are more readily translated by the selected host, e.g., thewell-known E. coli preference codons for E. coli expression.

Mutations can be introduced at particular loci by synthesizingoligonucleotides containing a mutant sequence, flanked by restrictionsites enabling ligation to fragments of the native sequence. Followingligation, the resulting reconstructed sequence encodes an analog havingthe desired amino acid insertion, substitution, or deletion.

Alternatively, oligonucleotide-directed site-specific mutagenesisprocedures can be employed to provide an altered gene having particularcodons altered according to the substitution, deletion, or insertionrequired. Exemplary methods of making the alterations set forth aboveare disclosed by Walder et al. (Gene 42:133, 1986); Bauer et al. (Gene37:73, 1985); Craik (BioTechniques, January 1985, 12-19); Smith et al.(Genetic Engineering: Principles and Methods, Plenum Press, 1981); andU.S. Pat. Nos. 4,518,584 and 4,737,462 disclose suitable techniques, andare incorporated by reference herein.

Due to code degeneracy, there can be considerable variation innucleotide sequences encoding the same amino acid sequence. Otherembodiments include sequences capable of hybridizing under moderatelystringent conditions (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mMEDTA (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight)to the DNA sequences encoding IL-17R, and other sequences which aredegenerate to those which encode the IL-17R. In a preferred embodiment,IL-17R analogs are at least about 70% identical in amino acid sequenceto the amino acid sequence of IL-17R proteins as set forth in SEQ IDNO:1 or SEQ ID NO:9. Similarly, analogs of IL-17R homologs are at leastabout 70% identical in amino acid sequence to the amino acid sequence ofthe native, homologous proteins. In a most preferred embodiment, analogsof IL-17R or homologs thereof are at least about 80% identical in aminoacid sequence to the native form of the inventive proteins.

Percent identity may be determined using a computer program, forexample, the GAP computer program described by Devereux et al. (Nucl.Acids Res. 12:387, 1984) and available from the University of WisconsinGenetics Computer Group (UWGCG). For fragments derived from the IL-17Rprotein, the identity is calculated based on that portion of the IL-17Rprotein that is present in the fragment. Similar methods can be used toanalyze homologs of IL-17R.

The ability of IL-17R analogs to bind CTLA-8 can be determined bytesting the ability of the analogs to inhibit IL-17 (CTLA-8) -induced Tcell proliferation. Alternatively, suitable assays, for example, anenzyme immunoassay or a dot blot, employing CTLA-8 or HSV13 (or ahomolog thereof which binds native IL-17R) can be used to assess theability of IL-17R analogs to bind CTLA-8. Such methods are well known inthe art.

The IL-17R proteins and analogs described herein will have numeroususes, including the preparation of pharmaceutical compositions. Theinventive proteins will also be useful in preparing kits that are usedto detect IL-17 or IL-17R, for example, in patient specimens. Such kitswill also find uses in detecting the interaction of IL-17 and IL-17R, asis necessary when screening for antagonists or mimetics of thisinteraction (for example, peptides or small molecules that inhibit ormimic, respectively, the interaction). A variety of assay formats areuseful in such kits, including (but not limited to) ELISA, dot blot,solid phase binding assays (such as those using a biosensor), rapidformat assays and bioassays.

Expression of Recombinant Receptors for IL-17

The proteins of the present invention are preferably produced byrecombinant DNA methods by inserting a DNA sequence encoding IL-17Rprotein or a homolog thereof into a recombinant expression vector andexpressing the DNA sequence in a recombinant microbial expression systemunder conditions promoting expression. DNA sequences encoding theproteins provided by this invention can be assembled from cDNA fragmentsand short oligonucleotide linkers, or from a series of oligonucleotides,to provide a synthetic gene which is capable of being inserted in arecombinant expression vector and expressed in a recombinanttranscriptional unit.

Recombinant expression vectors include synthetic or cDNA-derived DNAfragments encoding IL-17R, homologs, or bioequivalent analogs, operablylinked to suitable transcriptional or translational regulatory elementsderived from mammalian, microbial, viral or insect genes. Suchregulatory elements include a transcriptional promoter, an optionaloperator sequence to control transcription, a sequence encoding suitablemRNA ribosomal binding sites, and sequences which control thetermination of transcription and translation, as described in detailbelow. The ability to replicate in a host, usually conferred by anorigin of replication, and a selection gene to facilitate recognition oftransformants may additionally be incorporated.

DNA regions are operably linked when they are functionally related toeach other. For example, DNA for a signal peptide (secretory leader) isoperably linked to DNA for a polypeptide if it is expressed as aprecursor which participates in the secretion of the polypeptide; apromoter is operably linked to a coding sequence if it controls thetranscription of the sequence; or a ribosome binding site is operablylinked to a coding sequence if it is positioned so as to permittranslation. Generally, operably linked means contiguous and, in thecase of secretory leaders, contiguous and in reading frame. DNAsequences encoding IL-17R or homoloas which are to be expressed in amicroorganism will preferably contain no introns that could prematurelyterminate transcription of DNA into mRNA.

Useful expression vectors for bacterial use can comprise a selectablemarker and bacterial origin of replication derived from commerciallyavailable plasmids comprising genetic elements of the well known cloningvector pBR322 (ATCC 37017). Such commercial vectors include, forexample, pKK223-3 (Pharmacia Fine Chemicals, Uppsala, Sweden) and pGEM1(Promega Biotec, Madison, Wis., USA). These pBR322 "backbone" sectionsare combined with an appropriate promoter and the structural sequence tobe expressed. E. coli is typically transformed using derivatives ofpBR322, a plasmid derived from an E. coli species (Bolivar et al., Gene2:95, 1977). pBR322 contains genes for ampicillin and tetracyclineresistance and thus provides simple means for identifying transformedcells.

Promoters commonly used in recombinant microbial expression vectorsinclude the β-lactamase (penicillinase) and lactose promoter system(Chang et al., Nature 275:615, 1978; and Goeddel et al., Nature 281:544,1979), the tryptophan (trp) promoter system (Goeddel et al., Nucl. AcidsRes. 8:4057, 1980; and EPA 36,776) and tac promoter (Maniatis, MolecularCloning: A Laboratory Manual, Cold Spring Harbor Laboratory, p. 412,1982). A particularly useful bacterial expression system employs thephage λP_(L) promoter and cI857ts thermolabile repressor. Plasmnidvectors available from the American Type Culture Collection whichincorporate derivatives of the λ P_(L) promoter include plasmid pHUB2,resident in E. coli strain JMB9 (ATCC 37092) and pPLc28, resident in E.coli RR1 (ATCC 53082).

Suitable promoter sequences in yeast vectors include the promoters formetallothionein, 3-phosphoglycerate kinase (Hitzeman et al., J. Biol.Chem. 255:2073, 1980) or other glycolytic enzymes (Hess et al., J. Adv.Enzyme Reg. 7:149, 1968; and Holland et al., Biochem. 17:4900, 1978),such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase,pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphateisomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphateisomerase, phosphoglucose isomerase, and glucokinase. Suitable vectorsand promoters for use in yeast expression are further described in R.Hitzeman et al., EPA 73,657.

Preferred yeast vectors can be assembled using DNA sequences from pBR322for selection and replication in E. coli (Amp^(r) gene and origin ofreplication) and yeast DNA sequences including a glucose-repressibleADH2 promoter and α-factor secretion leader. The ADH2 promoter has beendescribed by Russell et al. (J. Biol. Chem. 258:2674, 1982) and Beier etal. (Nature 300:724, 1982). The yeast α-factor leader, which directssecretion of heterologous proteins, can be inserted between the promoterand the structural gene to be expressed. See, e.g., Kurjan et al., Cell30:933, 1982; and Bitter et al., Proc. Natl. Acad. Sci. USA 81:5330,1984. The leader sequence may be modified to contain, near its 3' end,one or more useful restriction sites to facilitate fusion of the leadersequence to foreign genes.

The transcriptional and translational control sequences in expressionvectors to be used in transforming vertebrate cells may be provided byviral sources. For example, commonly used promoters and enhancers arederived from Polyoma, Adenovirus 2, Simian Virus 40 (SV40), and humancytomegalovirus. DNA sequences derived from the SV40 viral genome, forexample, SV40 origin, early and late promoter, enhancer, splice, andpolyadenylation sites may be used to provide the other genetic elementsrequired for expression of a heterologous DNA sequence. The early andlate promoters are particularly useful because both are obtained easilyfrom the virus as a fragment which also contains the SV40 viral originof replication (Fiers et al., Nature 273:113, 1978). Smaller or largerSV40 fragments may also be used, provided the approximately 250 bpsequence extending from the Hind III site toward the BglI site locatedin the viral origin of replication is included. Further, viral genomicpromoter, control and/or signal sequences may be utilized, provided suchcontrol sequences are compatible with the host cell chosen. Exemplaryvectors can be constructed as disclosed by Okayama and Berg (Mol. Cell.Biol. 3:280, 1983).

A useful system for stable high level expression of mammalian receptorcDNAs in C127 murine mammary epithelial cells can be constructedsubstantially as described by Cosman et al. (Mol. Immunol. 23:935,1986). A preferred eukaryotic vector for expression of IL-17R DNA isreferred to as pDC406 (McMahan et al., EMBO J. 10:2821, 1991), andincludes regulatory sequences derived from SV40, human immunodeficiencyvirus (HIV), and Epstein-Barr virus (EBV). Other preferred vectorsinclude pDC409 and pDC410, which are derived from pDC406. pDC410 wasderived from pDC406 by substituting the EBV origin of replication withsequences encoding the SV40 large T antigen. pDC409 differs from pDC406in that a Bgl II restriction site outside of the multiple cloning sitehas been deleted, making the Bgl II site within the multiple cloningsite unique.

A useful cell line that allows for episomal replication of expressionvectors, such as pDC406 and pDC409, which contain the EBV origin ofreplication, is CV-1/EBNA (ATCC CRL 10478). The CV-1/EBNA cell line wasderived by transfection of the CV-1 cell line with a gene encodingEpstein-Barr virus nuclear antigen-1 (EBNA-1) and constitutively expressEBNA-1 driven from human CMV immediate-early enhancer/promoter.

Host Cells

Transformed host cells are cells which have been transformed ortransfected with expression vectors constructed using recombinant DNAtechniques and which contain sequences encoding the proteins of thepresent invention. Transformed host cells may express the desiredprotein (IL-17R or homologs thereof), but host cells transformed forpurposes of cloning or amplifying the inventive DNA do not need toexpress the protein. Expressed proteins will preferably be secreted intothe culture supernatant, depending on the DNA selected, but may bedeposited in the cell membrane.

Suitable host cells for expression of viral proteins includeprokaryotes, yeast or higher eukaryotic cells under the control ofappropriate promoters. Prokaryotes include gram negative or grampositive organisms, for example E. coli or Bacillus spp. Highereukaryotic cells include established cell lines of mammalian origin asdescribed below. Cell-free translation systems could also be employed toproduce viral proteins using RNAs derived from the DNA constructsdisclosed herein. Appropriate cloning and expression vectors for usewith bacterial, fungal, yeast, and mammalian cellular hosts aredescribed by Pouwels et al. (Cloning Vectors: A Laboratory Manual,Elsevier, N.Y., 1985), the relevant disclosure of which is herebyincorporated by reference.

Prokaryotic expression hosts may be used for expression of IL-17R orhomologs that do not require extensive proteolytic and disulfideprocessing. Prokaryotic expression vectors generally comprise one ormore phenotypic selectable markers, for example a gene encoding proteinsconferring antibiotic resistance or supplying an autotrophicrequirement, and an origin of replication recognized by the host toensure amplification within the host.

Suitable prokaryotic hosts for transformation include E. coli, Bacillussubtilis, Salmonella typhimurium, and various species within the generaPseudomonas, Streptomyces, and Staphylococcus, although others may alsobe employed as a matter of choice.

Recombinant IL-17R may also be expressed in yeast hosts, preferably fromthe Saccharomyces species, such as S. cerevisiae. Yeast of other genera,such as Pichia or Kluyveromyces may also be employed. Yeast vectors willgenerally contain an origin of replication from the 2μ yeast plasmid oran autonomously replicating sequence (ARS), promoter, DNA encoding theviral protein, sequences for polyadenylation and transcriptiontermination and a selection gene. Preferably, yeast vectors will includean origin of replication and selectable marker permitting transformationof both yeast and E. coli, e.g., the ampicillin resistance gene of E.coli and S. cerevisiae trp1 gene, which provides a selection marker fora mutant strain of yeast lacking the ability to grow in tryptophan, anda promoter derived from a highly expressed yeast gene to inducetranscription of a structural sequence downstream. The presence of thetrp1 lesion in the yeast host cell genome then provides an effectiveenvironment for detecting transformation by growth in the absence oftryptophan.

Suitable yeast transformation protocols are known to those of skill inthe art; an exemplary technique is described by Hinnen et al., Proc.Natl. Acad. Sci. USA 75:1929, 1978, selecting for Trp⁺ transformants ina selective medium consisting of 0.67% yeast nitrogen base, 0.5%casamino acids, 2% glucose, 10 μg/ml adenine and 20 μg/ml uracil. Hoststrains transformed by vectors comprising the ADH2 promoter may be grownfor expression in a rich medium consisting of 1% yeast extract, 2%peptone, and 1% glucose supplemented with 80 μg/ml adenine and 80 μg/mluracil. Derepression of the ADH2 promoter occurs upon exhaustion ofmedium glucose. Crude yeast supernatants are harvested by filtration andheld at 4° C. prior to further purification.

Various mammalian or insect cell culture systems can be employed toexpress recombinant protein. Baculovirus systems for production ofheterologous proteins in insect cells are reviewed by Luckow andSummers, Bio/Technology 6:47 (1988). Examples of suitable mammalian hostcell lines include the COS-7 lines of monkey kidney cells, described byGluzman (Cell 23:175, 1981), and other cell lines capable of expressingan appropriate vector including, for example, CV-1/EBNA (ATCC CRL10478), L cells, C127, 3T3, Chinese hamster ovary (CHO), HeLa and BHKcell lines. Mammalian expression vectors may comprise nontranscribedelements such as an origin of replication, a suitable promoter andenhancer linked to the gene to be expressed, and other 5' or 3' flankingnontranscribed sequences, and 5' or 3' nontranslated sequences, such asnecessary ribosome binding sites, a polyadenylation site, splice donorand acceptor sites, and transcriptional termination sequences.

Purification of Receptors for IL-17

Purified IL-17R, homologs, or analogs are prepared by culturing suitablehost/vector systems to express the recombinant translation products ofthe DNAs of the present invention, which are then purified from culturemedia or cell extracts. For example, supernatants from systems whichsecrete recombinant protein into culture media can be first concentratedusing a commercially available protein concentration filter, forexample, an Amicon or Millipore Pellicon ultrafiltration unit.

Following the concentration step, the concentrate can be applied to asuitable purification matrix. For example, a suitable affinity matrixcan comprise a counter structure protein or lectin or antibody moleculebound to a suitable support. Alternatively, an anion exchange resin canbe employed, for example, a matrix or substrate having pendantdiethylaminoethyl (DEAE) groups. The matrices can be acrylamide,agarose, dextran, cellulose or other types commonly employed in proteinpurification. Alternatively, a cation exchange step can be employed.Suitable cation exchangers include various insoluble matrices comprisingsulfopropyl or carboxymethyl groups. Sulfopropyl groups are preferred.Gel filtration chromatography also provides a means of purifying theinventive proteins.

Affinity chromatography is a particularly preferred method of purifyingIL-17R and homologs thereof. For example, a IL-17R expressed as a fusionprotein comprising an immunoglobulin Fc region can be purified usingProtein A or Protein G affinity chromatography. Moreover, a IL-17Rprotein comprising an oligomerizing zipper domain may be purified on aresin comprising an antibody specific to the oligomerizing zipperdomain. Monoclonal antibodies against the IL-17R protein may also beuseful in affinity chromatography purification, by utilizing methodsthat are well-known in the art. A ligand (i.e., IL-17 or HVS-13) mayalso be used to prepare an affinity matrix for affinity purification ofIL-17R.

Finally, one or more reversed-phase high performance liquidchromatography (RP-HPLC) steps employing hydrophobic RP-HPLC media,e.g., silica gel having pendant methyl or other aliphatic groups, can beemployed to further purify a IL-17R composition. Some or all of theforegoing purification steps, in various combinations, can also beemployed to provide a homogeneous recombinant protein.

Recombinant protein produced in bacterial culture is usually isolated byinitial extraction from cell pellets, followed by one or moreconcentration, salting-out, aqueous ion exchange or size exclusionchromatography steps. Finally, high performance liquid chromatography(HPLC) can be employed for final purification steps. Microbial cellsemployed in expression of recombinant viral protein can be disrupted byany convenient method, including freeze-thaw cycling, sonication,mechanical disruption, or use of cell lysing agents.

Fermentation of yeast which express the inventive protein as a secretedprotein greatly simplifies purification. Secreted recombinant proteinresulting from a large-scale fermentation can be purified by methodsanalogous to those disclosed by Urdal et al. (J. Chromatog. 296:171,1984). This reference describes two sequential, reversed-phase HPLCsteps for purification of recombinant human GM-CSF on a preparative HPLCcolumn.

Protein synthesized in recombinant culture is characterized by thepresence of cell components, including proteins, in amounts and of acharacter which depend upon the purification steps taken to recover theinventive protein from the culture. These components ordinarily will beof yeast, prokaryotic or non-human higher eukaryotic origin andpreferably are present in innocuous contaminant quantities, on the orderof less than about 1 percent by weight. Further, recombinant cellculture enables the production of the inventive proteins free of otherproteins which may be normally associated with the proteins as they arefound in nature in the species of origin.

Administration of IL-17R Compositions

The present invention provides methods of using therapeutic compositionscomprising an effective amount of a protein and a suitable diluent andcarrier, and methods for regulating an immune response. The use ofIL-17R or homologs in conjunction with soluble cytokine receptors orcytokines, or other immunoregulatory molecules is also contemplated.Moreover, DNA encoding soluble IL-17R will also be useful; a tissue ororgan to be transplanted can be transfected with the DNA by any methodknown in the art. The organ or tissue thus expresses soluble IL-17R,which acts in the localized area of the graft to suppress rejection ofthe graft. Similar methods comprising administering such DNA's to thesite of the graft will also show efficacy in ameliorating graftrejection.

For therapeutic use, purified protein is administered to a patient,preferably a human, for treatment in a manner appropriate to theindication. Thus, for example, IL-17R protein compositions administeredto regulate immune function can be given by bolus injection, continuousinfusion, sustained release from implants, or other suitable technique.Typically, a therapeutic agent will be administered in the form of acomposition comprising purified IL-17R, in conjunction withphysiologically acceptable carriers, excipients or diluents. Suchcarriers will be nontoxic to recipients at the dosages andconcentrations employed.

Ordinarily, the preparation of such protein compositions entailscombining the inventive protein with buffers, antioxidants such asascorbic acid, low molecular weight (less than about 10 residues)polypeptides, proteins, amino acids, carbohydrates including glucose,sucrose or dextrins, chelating agents such as EDTA, glutathione andother stabilizers and excipients. Neutral buffered saline or salinemixed with conspecific serum albumin are exemplary appropriate diluents.Preferably, product is formulated as a lyophilizate using appropriateexcipient solutions (e.g., sucrose) as diluents. Appropriate dosages canbe determined in trials. The amount and frequency of administration willdepend, of course, on such factors as the nature and severity of theindication being treated, the desired response, the condition of thepatient, and so forth.

Receptors for IL-17 (CTLA-8) can be administered for the purpose ofinhibiting T cell proliferation, or for inhibiting T cell activation.Soluble IL-17R are thus likely to be useful in preventing or treatingorgan or graft rejection, autoimmune disease, allergy or asthma. Theinventive receptor proteins will also be useful for prevention ortreatment of inflammatory disease in which activated T cells play arole. Similarly, HVS13 and homologs thereof stimulate B cellproliferation and immunoglobulin secretion; thus, receptors that bindHVS13 or CTLA-8 will be useful in vivo to inhibit B cell proliferationor immunoglobulin secretion. Receptors for CTLA-8 will also be useful toinhibit the binding of HVS13 or CTLA-8 to cells expressing IL-17R.

The following examples are offered by way of illustration, and not byway of limitation. Those skilled in the art will recognize thatvariations of the invention embodied in the examples can be made,especially in light of the teachings of the various references citedherein, the disclosures of which are incorporated by reference.

EXAMPLE 1

This example describes identification of cells that express a receptor(or counterstructure) for HVS13/mCTLA8. A chimeric protein (HVS13 typeII Fc) consisting of an Fc region of a human immunoglobulin (SEQ IDNO:4) followed by the amino acid 19 to 151 of HVS13 (SEQ ID NO:8) wasprepared. A murine CTLA8/Fc (mCTLA8/Fc) was constructed by fusing aminoacid 22 to 150 of mCTLA8 (SEQ ID NO:6) to the Fc region of human IgG1. Acontrol Fc protein was constructed by a similar method. The HVS13/Fc andmCTLA-8 proteins were expressed and used to identify cell sources byflow cytometry.

Cells (1×10⁶) were preincubated on ice for 30 minutes in 100 μl of FACSbuffer (PBS, 1% FCS and 0.1% NaN3) containing 2% normal goat serum and2% normal rabbit serum to block nonspecific binding. 100 μl of HVS13/Fc,mCTLA-8/Fc or control/Fc protein was added at 5 μg/ml and incubated onice for 30 min. After washing, the cells were stained with biotinlabeled anti human IgG (Fc specific) followed by PE-conjugatedstreptavidin (Becton Dickson & Co, Mountain View, Calif.) in 100 μl ofFACS buffer. Cells were then washed and analyzed using a FACScan (BectonDickinson). A minimum of 5,000 cells were analyzed for each sample. Morethan a dozen cell lines were screened and it was found that bothHVS13/Fc and mCTLA8/Fc fusion proteins bound specifically to the murinethymoma cell line EL4. These cells did not bind to the control/Fc fusionprotein.

EXAMPLE 2

This example describes cloning of the gene that encodes IL-17R. Afteridentification of a source for HVS13 counterstructure, an EL4 mammalianexpression library was screened by a slide-binding autoradiographicmethod (Gearing et al., EMBO J. 8:3667, 1989). CV1/EBNA cells weremaintained in Dulbecco's modified Eagle's medium (DMEM) containing 10%(v/v) fetal calf serum (FCS) at 37° C. in a humidified atmospherecontaining 10% CO2 and passaged twice weekly. Subconfluent CV1/EBNA cellmonolayers on fibronectin-treated chamber slides (Labtek) weretransfected by a chloroquine-mediated DEAE-dextran procedure withplasmid DNAs derived from pooled transformants (2,000 transformants perpool) of murine EL4 cDNA library.

The CV1/EBNA cells transfected with the murine EL4 cDNA pools wereassayed for HVS13/Fc binding two days after transfection using [¹²⁵ I]labeled goat anti-human IgG binding and slide autoradiography.Transfected cell monolayers were washed with binding medium (RPMI 1640containing 1% bovine serum albumin and 50 mg/ml non-fat dry milk), thenincubated with 1 μg/ml of HVS13/Fc for one hour at room temperature.Cells were washed, incubated with ¹²⁵ I-labeled goat anti-human IgG (NewEngland nuclear, Cambridge, Mass.). Cells were washed twice with bindingmedium, three times with PBS, and fixed in PBS containing 2.5%gluteraldehyde for 30 minutes, washed twice more with PBS and air dried.The chamber slides were then dipped in Kodak GTNB-2 photographicemulsion and exposed for 3 days at 4° C. before developing.

Forty pools of approximately 2,000 cDNA each were transfected intoCV1/EBNA cells. Two pools of cDNA were found to confer binding toHVS13/Fc protein. These pools were broken down to pools of 100 cDNAs,and subsequently to individual clones. Two single cDNA clones wereisolated. These clones were transfected into CV1/EBNA to determinewhether the protein encoded thereby conferred binding to both HVS13/Fcand mCTLA8/Fc. Both HVS/Fc and mCTLA8/Fc bound to CV1/EBNA cellstransfected with the cloned cDNA, but not to cells transfected withempty vector. Control/Fc did not bind to either of them.

Sequencing of these clones found that they contained a 3.2 kb and 1.7 kbinsert derived from same mRNA. The 3.2 kb clone contained an openreading frame of 2595 bp surrounded by 120 bp at the 5' noncodingsequence and 573 bp of 3' noncoding sequence. There were no in-framestop codons upstream of the predicted initiator methionine, which ispreceded by a purine residue (guanine) at -3 position, the mostimportant indicator of a good translation initiation site (Kozak, Mol.Cell. Biol. 9:5134, 1989). It also has a guanine at +4 position, makingit an optimal for translation initiation. The open reading frame ispredicted to encode a type I transmembrane protein of 864 amino acids.The nucleotide and predicted amino acid sequence is shown in SEQ IDNOs:1 and 2.

Computer analysis indicated that the protein has an N-terminal signalpeptide with a cleavage site between amino acid 31 and 32. The signalpeptide is followed by a 291 amino acid extracellular domain, a 21 aminoacid transmembrane domain, and a 521 amino acid cytoplasmic tail. Thereare eight potential N-linked glycosylation sites in the extracellulardomain of the protein. The predicted molecular weight for this proteinis 97.8 kilodaltons with an estimated isoelectric point of 4.85.Comparison of both nucleotide and amino acid sequences with the GenBankor EMBL databases found no significant homology with known nucleotideand protein sequences.

In order to determine the cellular and tissue distribution of IL-17RmRNA, poly (A)⁺ RNA derived from various murine cell lines or tissueswas examined by Northern blot analysis using the IL-17R cDNA as a probe.Filters containing poly(A)⁺ RNA (2 μg per lane) from various tissueswere purchased from Clontech (Palo Alto, Calif.). Polyadenylated RNAfrom various cells or cell lines were isolated, fractionated (2 μg perlane) on a 1% agarose formaldehyde gel, blotted onto Hybond nylonmembrane (Amersham). Filters were probed with an anti-sense RNAriboprobe corresponding to the coding region of IL-17R cDNA.Hybridization was performed at 63° C. followed by three washings in0.2%×SSC, 0.1% SDS at 68° C. Blots were exposed for 8 to 48 hr at -70°C.

The IL-17R probe hybrid ized to a single species oft RNA of approximately 3.7 kb in all tissues. Among the tissues examined, stronghybridizing signals were observed in spleen and kidney. Moderate signalswere observed in lung and liver, and weaker signals in brain, heart,skeletal muscle and testes. Similar size mRNAs were detected in thefollowing cells and cell lines: fetal liver epithelial cells (D11),fibroblast (3T3), rat intestinal epithelial cells (1CE6), splenic Bcells, muscle cells (BB4), mast cells (H7), triple negative thymus cells(TN), pre-B cells (70Z/3), T cell hybridoma (EM); and T cell clones 7C2and D10. All the cell lines tested were found to express IL-17R mRNA,suggesting a ubiquitous expression of IL-17R message.

EXAMPLE 3

This example describes construction of a construct to express a solubleIL-17R/Flag® protein referred to as IL-17R/Flag. IL-17R/Flag® contains aleader sequence, and the region of IL-17R from amino acid 1 to aminoacid 322 (SEQ ID NO:1), and the octapeptide referred to as Flag® (SEQ IDNO:3). The construct is prepared essentially as described for othersoluble constructs, by ligating a DNA fragment encoding amino acids 1througah 322 of SEQ ID NO:1 (prepared as described in Example 4) into anappropriate expression vector which contains a suitable leader sequence.The resultant DNA construct is transfected into a suitable cell line.such as the monkey kidney cell line CV-1/EBNA (ATCC CRL 10478).IL-f7R/Flag® may be purified using a Flag® antibody affinity column, andanalyzed for biological activity using any of the methods describedherein.

EXAMPLE 4

This example describes construction of a IL-17R DNA construct to expressa IL-17R/Fc fusion protein. A soluble form of IL-17R fused to the Fcregion of human IgG1 was constructed in the mammalian expression vectorpDC409 in the following way: A pair of oligonucleotide primerscontaining a sense sequence and an antisense sequence of IL-17R weresynthesized. The sense primer contained a Sal I site at the 5' end ofthe cDNA and antisense primer contained a Bgl II site and contained theIL-17R truncated just before the transmembrane region and a stop codon.A 980 bp DNA framgent was amplified from IL-17R cDNA. The PCR productwas cut with Sal I and Bgl II and used in a three way ligation with afragment carrying the human IgG1 region cut with Bgl II and Not I into aplasmid (pDC49; see U.S. Ser. No. 08/235,397) previously cut with Sal Iand Not N. The encoded insert contained the nucleotides encoding theamino acid sequence of residues 1 to 322 of IL-17R (SEQ ID NO:1). Thesequence was confirmed by sequaencind the whole, region.

The IL-l7R/Fc expression plasmids were transfected into CV-1/EBNA cells,and supenatants were collected for 1 week. The CTLA-8/Fc fusion proteinswere purified on a protein A sepharose column (Pharmacia, Uppsala,Sweden) as described below, Protein concentration was deterdmbined by anenzyme-linked immunoadsorbent assay specific for the constant domain ofhuman IgG1 and by BCA analysis (Pharmacia), and purity was confirmed bySDS-polyacrylalide gel electrophoresis analysis followed by silver stainof the gel.

EXAMPLE 5

This example describes purification of IL-17R fusion proteins. IL-17R/Fcfusion protein is purified by conventional methods using Protein A orProtein G chromatography. Approximately one liter of culture supernatantcontaining IL-l7R/Fc fusion protein is purified by filtering mammaliancell supernatants (e.g., in a 0.45 m filter) and applying filtrate to aprotein A/G antibody affinity column (Schleicher and Schuell, Keene, N.H.) at 4° C. at a flow rate of 80 ml/hr for a 1.5 cm×12.0 cm column. Thecolumn is washed with 0.5 M NaCl in PBS until free protein is notdetected in the wash buffer. Finally, the column is washed with PBS.Bound fusion protein is eluted from the column with 25 mM citratebuffer, pH 2.8, and brought to pH 7 with 500 mM Hepes buffer, pH 9.1.

A IL-17R fusion protein comprising Flag® may also be detected and/orpurified using an antibody that binds Flag®, substantially as describedin Hopp et al., Bio/Technology 6:1204 (1988). Biological activity ismeasured by inhibition of CTLA-8 activity in any biological assay whichquantifies the co-stimulatory effect of CTLA-8, for example, asdescribed in the Examples herein.

EXAMPLE 6

This example illustrates the preparation of monoclonal antibodiesagainst IL-17R. Preparations of purified recombinant IL-17R, forexample, or transfected cells expressing high levels of IL-17R, areemployed to generate monoclonal antibodies against IL-17R usingconventional techniques, such as those disclosed in U.S. Pat. No.4,411,993. Such antibodies are likely to be useful in interfering withIL-17R binding to CTLA-8, as components of diagnostic or research assaysfor IL-17R, or in affinity purification of IL-17R.

To immunize rodents, IL-17R immunogen is emulsified in an adjuvant (suchas complete or incomplete Freund's adjuvant, alum, or another adjuvant,such as Ribi adjuvant R700 (Ribi, Hamilton, Mont.), and injected inamounts ranging from 10-100 μg subcutaneously into a selected rodent,for example, BALB/c mice or Lewis rats. Ten days to three weeks dayslater, the immunized animals are boosted with additional immunogen andperiodically boosted thereafter on a weekly, biweekly or every thirdweek immunization schedule. Serum samples are periodically taken byretro-orbital bleeding or tail-tip excision for testing by dot-blotassay (antibody sandwich), ELISA (enzyme-linked immunosorbent assay),immunoprecipitation, or other suitable assays, including FACS analysis.Following detection of an appropriate antibody titer, positive animalsare given an intravenous injection of antigen in saline. Three to fourdays later, the animals are sacrificed, splenocytes harvested, and fusedto a murine myeloma cell line (e.g., NS1 or preferably Ag 8.653 [ATCCCRL 1580]). Hybridoma cell lines generated by this procedure are platedin multiple microtiter plates in a selective medium (for example, onecontaining hypoxanthine, aminopterin, and thymidine, or HAT) to inhibitproliferation of non-fused cells, myeloma-myeloma hybrids, andsplenocyte-splenocyte hybrids.

Hybridoma clones thus generated can be screened by ELISA for reactivitywith IL-17R, for example, by adaptations of the techniques disclosed byEngvall et al., Immitnochem. 8:871 (1971) and in U.S. Pat. No.4,703,004. A preferred screening technique is the antibody capturetechnique described by Beckman et al., J. Immunol. 144:4212 (1990).Positive clones are then injected into the peritoneal cavities ofsyngeneic rodents to produce ascites containing high concentrations (>1mg/ml) of anti-IL-17R monoclonal antibody. The resulting monoclonalantibody can be purified by ammonium sulfate precipitation followed bygel exclusion chromatography. Alternatively, affinity chromatographybased upon binding of antibody to protein A or protein G can also beused, as can affinity chromatography based upon binding to IL-17Rprotein.

EXAMPLE 7

This example illustrates the ability of IL-17R to inhibit theproliferative response of T cells to mitogens. Lymphoid organs wereharvested aseptically and cell suspension was created. Splenic and lymphnode T cells were isolated from the cell suspension. The purity of theresulting splenic T cell preparations was routinely >95% CD3⁺ and <1%sIgM⁺. Purified murine splenic T cells (2×10⁵ /well) were cultured witheither 1% PHA or 1 μg/ml Con A, and a soluble IL-17R was titered intothe assay. Proliferation was determined after 3 days with the additionof 1 μCi [³ H]thyrnidine. Secretion of cytokines (Interleukin-2) wasdetermined for murine T cells cultured for 24 hr with 1 μg/ml of Con Ain the presence or absence of 10 μg/ml of IL-17R.Fc or in the presenceof a control Fc protein. IL-2 production was measured by ELISA andresults expressed as ng/ml IL-2 produced.

Soluble IL-17R/Fc significantly inhibited the mitogen-inducedproliferation of purified murine splenic T cells in a dose dependentmanner, while a control Fc had no effect on the murine T cellproliferation. Complete inhibition of mitogen induced proliferation wasobserved at a soluble IL-17R.Fc concentration of 10 μg/ml. Analysis ofIL-2 production by splenic T cells activated with Con A in the presenceor absence of IL-17R.Fc in the culture revealed that addition ofIL-17R.Fc to the T-cell culture inhibited IL-2 production to levels8-9-fold lower than those observed in cultures containing media alone ormedia plus a control Fc protein. Similar results were observed whenpurified human T cells were used.

EXAMPLE 8

This example presents the isolation of a DNA encoding human IL-17R bycross species hybridization. A human peripheral blood lymphocyte librarywas prepared and screened substantially as described in U.S. Ser. No.08/249,189, using murine IL-17R DNA under moderately high stringencyconditions. Several clones of varying length were obtained. Sequencingdata indicated that the human IL-17R was approximately 76% identical tomurine IL-17R at the nucleotide level. The nucleotide and predictedamino acid sequence of human IL-17R is shown in SEQ ID NOs:10 and 11. Aplasmid (pGEMBL) containing DNA encoding the human IL-17 receptor(referred to as pGEMBL-HuIL-17R) in E. coli DH10, was deposited with theAmerican Type Culture Collection, 12301 Parklawn Drive, Rockville, Md.20852-1776, USA, on Jun. 5, 1995, under the conditions of the BudapestTreaty, and assigned accession number 69834.

The human IL-17R shared many features with the murine IL-17 R. Computeranalysis indicated that the protein has an N-terminal signal peptidewith a cleavage site between amino acid 27 and 28. The signal peptide isfollowed by a 293 amino acid extracellular domain, a 21 amino acidtransmembrane domain, and a 525 amino acid cytoplasmic tail. SolubleIL-17R comprises the signal peptide and the extracellular domain(residues 1 to 320 of SEQ ID NO:1) or a fragment thereof. Alternatively,a different signal peptide can be substituted for the native signalpeptide. A Type I Fc fusion protein (wherein DNA encoding the Fc regionof an immunoglobulin molecule is fused to DNA encoding the IL-17Rimmediately before, and in place of, the DNA encoding the transmembraneregion of the IL-17R) was prepared, substantially as described inExample 4. A soluble hIL-17R protein can be also expressed substantiallyas described in Example 3, or by any other method of preparing andexpressing the extracellular domain of IL-17R or a fragment thereof.

EXAMPLE 9

This example presents the localization and fine mapping of the murineIL-17R gene. A panel of DNA samples from an interspecific cross that hasbeen characterized for over 900 genetic markers throughout the genomewas analyzed. The genetic markers included in this map span between 50and 80 centi-Morgans on each mouse autosome and the X chromosome (Chr)(Saunders and Seldin, Genomics 8:524, 1990; Watson et al., MammalianGenome 2:158, 1992).

Initially, DNA from the two parental mice [C3H/HeJ-gld and (C3H/HeJ-gldx Mus spretus) F1] were digested with various restriction endonucleasesand hybridized with the IL-17R cDNA probe to determine restrictionfragment length variants (RFLVs) to allow haplotype analyses.Informative Bgl1 RFLVs were detected: C3H/HeJ-gld, 10.0 kb; Mus spretus,7.8 kb and 2.2 kb). In each of the backcross mice either the C3H/HeJ-gldparental band or all three bands (both Mus spretus bands and a halfintensity C3H/HEJ-gld band) were observed indicating that a single locuswas detected.

Comparison of the haplotype distribution of the IL-17R RFLVs indicatedthat this gene cosegregated in 111 of the 114 meiotic events examinedwith the Raf1 gene locus on mouse Chr 6. The best gene order (Bishop,Genet. Epidemiol. 2:349, 1985) ±the standard deviation (Green, InGenetics and Probability in Animal Breeding Experiments. E. Green, ed.;Macmillan, New York, pp. 77-113, 1981) was: (centromere) Raf1-2.6 cM±1.5 cM-IL-17R-2.5 cM±1.5cM-Cd4.

EXAMPLE 10

This example demonstrates that soluble IL-17R suppresses rejection oforgan grafts in vivo. Hearts from neonatal C57BL/6 (H-2^(b)) mice (lessthan 24 hours old) were transplanted into the ear pinnae of adult BALB/c(H-2^(d)) recipients substantially as described in U.S. Pat. No.5,492,888, issued Feb. 20, 1996 (utilizing the method of Fulmer et al.,Am. J. Anat. 113:273, 1963, modified as described by Trager et al.,Transplantation 47:587, 1989, and Van Buren et al., Transplant. Proc.15:2967, 1983). Survival of the transplanted hearts was assessed byvisually inspecting the grafts for pulsatile activity, as determined byexamining the ear-heart grafts of anesthetized recipients under adissecting microscope with soft reflected light beginning on day 5 or 6post transplant. The time of graft rejection was defined as the dayafter transplantation on which contractile activity ceased.

In one set of experiments, neonatal hearts were removed, rinsed withsterile PBS to remove excess blood, and placed into prepared ear pinnae.Recipient mice were given either soluble murine IL-17R/Fc (100 μg in 200μl; see Example 4 herein) or rat IgG as a control, i.p. on days 0through 3 post transplantation. In a second set of experiments, therecipient mice were injected with IL-17R or human IgG on days 0, 1 and2; the quantity and route of injection were as done previously. Theresults of these experiments are shown in Table 1.

                  TABLE 1                                                         ______________________________________                                        Effects of Soluble Murine IL-17R (smuIL-17R) on                               Neovascularized Heterotopic Cardiac Allograft Survival                               Treatment                                                                             Survival   Median Survival Time ±                                  Group   Time (days)                                                                              S. D.                                               ______________________________________                                        Experiment 1                                                                           rat IgG   11, 14, 14, 14                                                                           13 ± 1.5                                              smuIL-17R 19, 19, 19, 21                                                                           20 ± 1.0                                     Experiment 2                                                                           human IgG 13, 13, 13, 15                                                                           14 ± 1.0                                              smuIL-17R 20, 20, 20, 20                                                                           20 ± 0.0                                     ______________________________________                                    

Table 1 shows that heart allografts survived approximately 13 days inindividual control mice treated with rat IgG. When allograft recipientswere given up to four daily injections of soluble IL-17R, graft survivalwas prolonged, with a median survival of 20, approximately seven dayslonger than the survival time of identical grafts in control mice. Whena prolonged release of the IL-17R was obtained by encapsulating thesoluble IL-17R in alginate beads, it was observed that a sin-leadministration of 100 μg soluble IL-17R prolonged graft s urvival inmuch the same manner as observed previously with soluble IL-17R insolution. These results demonstrate that soluble IL-17R suppressesrejection of grafted tissues.

EXAMPLE 11

This example demonstrates that DNA encoding soluble IL-17R will beuseful in suppressing rejection of organ grafts in vivo. Hearts fromneonatal C57BL/6 (H-2^(b)) mice were transplanted into the ear pinnae ofadult BALB/c (H-2^(d)) recipients as described in Example 10 above,except that the hearts were injected with 15 μl of PBS containing eitherIL-17R/Fc-encoding, DNA (pDC409-IL-17R; Example 4) or control DNA (emptypDC409) at a concentration of about 1 mg/ml, into a ventricle. A 30gauge needle was used, and care was taken to minimize trauma to theheart. The transfected hearts were then transplanted into BALB/crecipients and graft survival determined as described previously.Results are presented below in Table 2.

                  TABLE 2                                                         ______________________________________                                        Effects of Expression of Soluble Murine IL-17R by Cardiac Cells               on Neovascularized Heterotopic Cardiac Allograft Survival                     Treatment                Median Survival Time ±                            Group      Survival Time (days)                                                                        S. D.                                                ______________________________________                                        rat IgG    13, 15, 15, 15, 18                                                                          15 ± 1.8                                          smuIL-17R  20, 25, 28, >60, >60                                                                        ND*                                                  ______________________________________                                         *ND: Not done; median survival time could not be calculated since two mic     still show pulsatile grafts more than two months after transplantation.  

Table 2 shows that heart allografts survived approximately 15 days inindividual control mice transplanted with hearts transfected with emptyvector. When the transplanted hearts were transfected with DNA encodingsoluble IL-17R, graft survival was prolonged. For three of the five micein this group, grafts survived on average approximately 24 days, ninedays longer than the survival time of identical grafts in control mice.The grafts given the other two mice were still puslatile (i.e., had notbeen rejected) more than 60 days post transplant., and had apparentlybeen accepted by the recipients. These results demonstrate thattransfecting tissues to be grafted with DNA encoding soluble IL-17Rameliorates rejection of those tissues by the recipient.

    __________________________________________________________________________    #             SEQUENCE LISTING                                                - (1) GENERAL INFORMATION:                                                    -    (iii) NUMBER OF SEQUENCES: 10                                            - (2) INFORMATION FOR SEQ ID NO:1:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 3288 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA to mRNA                                        -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Mouse                                                           (B) STRAIN: HVS13 recep - #tor                                      -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 121..2715                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                 - GTCGACTGGA ACGAGACGAC CTGCTGCCGA CGAGCGCCAG TCCTCGGCCG GG - #AAAGCCAT         60                                                                          - CGCGGGCCCT CGCTGTCGCG CGGAGCCAGC TGCGAGCGCT CCGCGACCGG GC - #CGAGGGCT        120                                                                          - ATG GCG ATT CGG CGC TGC TGG CCA CGG GTC GT - #C CCC GGG CCC GCG CTG          168                                                                          Met Ala Ile Arg Arg Cys Trp Pro Arg Val Va - #l Pro Gly Pro Ala Leu           #                 15                                                          - GGA TGG CTG CTT CTG CTG CTG AAC GTT CTG GC - #C CCG GGC CGC GCC TCC          216                                                                          Gly Trp Leu Leu Leu Leu Leu Asn Val Leu Al - #a Pro Gly Arg Ala Ser           #             30                                                              - CCG CGC CTC CTC GAC TTC CCG GCT CCG GTC TG - #C GCG CAG GAG GGG CTG          264                                                                          Pro Arg Leu Leu Asp Phe Pro Ala Pro Val Cy - #s Ala Gln Glu Gly Leu           #         45                                                                  - AGC TGC AGA GTC AAG AAT AGT ACT TGT CTG GA - #T GAC AGC TGG ATC CAC          312                                                                          Ser Cys Arg Val Lys Asn Ser Thr Cys Leu As - #p Asp Ser Trp Ile His           #     60                                                                      - CCC AAA AAC CTG ACC CCG TCT TCC CCA AAA AA - #C ATC TAT ATC AAT CTT          360                                                                          Pro Lys Asn Leu Thr Pro Ser Ser Pro Lys As - #n Ile Tyr Ile Asn Leu           # 80                                                                          - AGT GTT TCC TCT ACC CAG CAC GGA GAA TTA GT - #C CCT GTG TTG CAT GTT          408                                                                          Ser Val Ser Ser Thr Gln His Gly Glu Leu Va - #l Pro Val Leu His Val           #                 95                                                          - GAG TGG ACC CTG CAG ACA GAT GCC AGC ATC CT - #G TAC CTC GAG GGT GCA          456                                                                          Glu Trp Thr Leu Gln Thr Asp Ala Ser Ile Le - #u Tyr Leu Glu Gly Ala           #           110                                                               - GAG CTG TCC GTC CTG CAG CTG AAC ACC AAT GA - #G CGG CTG TGT GTC AAG          504                                                                          Glu Leu Ser Val Leu Gln Leu Asn Thr Asn Gl - #u Arg Leu Cys Val Lys           #       125                                                                   - TTC CAG TTT CTG TCC ATG CTG CAG CAT CAC CG - #T AAG CGG TGG CGG TTT          552                                                                          Phe Gln Phe Leu Ser Met Leu Gln His His Ar - #g Lys Arg Trp Arg Phe           #   140                                                                       - TCC TTC AGC CAC TTT GTG GTA GAT CCT GGC CA - #G GAG TAT GAA GTG ACT          600                                                                          Ser Phe Ser His Phe Val Val Asp Pro Gly Gl - #n Glu Tyr Glu Val Thr           145                 1 - #50                 1 - #55                 1 -       #60                                                                           - GTT CAC CAC CTG CCG AAG CCC ATC CCT GAT GG - #G GAC CCA AAC CAC AAA          648                                                                          Val His His Leu Pro Lys Pro Ile Pro Asp Gl - #y Asp Pro Asn His Lys           #               175                                                           - TCC AAG ATC ATC TTT GTG CCT GAC TGT GAG GA - #C AGC AAG ATG AAG ATG          696                                                                          Ser Lys Ile Ile Phe Val Pro Asp Cys Glu As - #p Ser Lys Met Lys Met           #           190                                                               - ACT ACC TCA TGC GTG AGC TCA GGC AGC CTT TG - #G GAT CCC AAC ATC ACT          744                                                                          Thr Thr Ser Cys Val Ser Ser Gly Ser Leu Tr - #p Asp Pro Asn Ile Thr           #       205                                                                   - GTG GAG ACC TTG GAC ACA CAG CAT CTG CGA GT - #G GAC TTC ACC CTG TGG          792                                                                          Val Glu Thr Leu Asp Thr Gln His Leu Arg Va - #l Asp Phe Thr Leu Trp           #   220                                                                       - AAT GAA TCC ACC CCC TAC CAG GTC CTG CTG GA - #A AGT TTC TCC GAC TCA          840                                                                          Asn Glu Ser Thr Pro Tyr Gln Val Leu Leu Gl - #u Ser Phe Ser Asp Ser           225                 2 - #30                 2 - #35                 2 -       #40                                                                           - GAG AAC CAC AGC TGC TTT GAT GTC GTT AAA CA - #A ATA TTT GCG CCC AGG          888                                                                          Glu Asn His Ser Cys Phe Asp Val Val Lys Gl - #n Ile Phe Ala Pro Arg           #               255                                                           - CAA GAA GAA TTC CAT CAG CGA GCT AAT GTC AC - #A TTC ACT CTA AGC AAG          936                                                                          Gln Glu Glu Phe His Gln Arg Ala Asn Val Th - #r Phe Thr Leu Ser Lys           #           270                                                               - TTT CAC TGG TGC TGC CAT CAC CAC GTG CAG GT - #C CAG CCC TTC TTC AGC          984                                                                          Phe His Trp Cys Cys His His His Val Gln Va - #l Gln Pro Phe Phe Ser           #       285                                                                   - AGC TGC CTA AAT GAC TGT TTG AGA CAC GCT GT - #G ACT GTG CCC TGC CCA         1032                                                                          Ser Cys Leu Asn Asp Cys Leu Arg His Ala Va - #l Thr Val Pro Cys Pro           #   300                                                                       - GTA ATC TCA AAT ACC ACA GTT CCC AAG CCA GT - #T GCA GAC TAC ATT CCC         1080                                                                          Val Ile Ser Asn Thr Thr Val Pro Lys Pro Va - #l Ala Asp Tyr Ile Pro           305                 3 - #10                 3 - #15                 3 -       #20                                                                           - CTG TGG GTG TAT GGC CTC ATC ACA CTC ATC GC - #C ATT CTG CTG GTG GGA         1128                                                                          Leu Trp Val Tyr Gly Leu Ile Thr Leu Ile Al - #a Ile Leu Leu Val Gly           #               335                                                           - TCT GTC ATC GTG CTG ATC ATC TGT ATG ACC TG - #G AGG CTT TCT GGC GCC         1176                                                                          Ser Val Ile Val Leu Ile Ile Cys Met Thr Tr - #p Arg Leu Ser Gly Ala           #           350                                                               - GAT CAA GAG AAA CAT GGT GAT GAC TCC AAA AT - #C AAT GGC ATC TTG CCC         1224                                                                          Asp Gln Glu Lys His Gly Asp Asp Ser Lys Il - #e Asn Gly Ile Leu Pro           #       365                                                                   - GTA GCA GAC CTG ACT CCC CCA CCC CTG AGG CC - #C AGG AAG GTC TGG ATC         1272                                                                          Val Ala Asp Leu Thr Pro Pro Pro Leu Arg Pr - #o Arg Lys Val Trp Ile           #   380                                                                       - GTC TAC TCG GCC GAC CAC CCC CTC TAT GTG GA - #G GTG GTC CTA AAG TTC         1320                                                                          Val Tyr Ser Ala Asp His Pro Leu Tyr Val Gl - #u Val Val Leu Lys Phe           385                 3 - #90                 3 - #95                 4 -       #00                                                                           - GCC CAG TTC CTG ATC ACT GCC TGT GGC ACT GA - #A GTA GCC CTT GAC CTC         1368                                                                          Ala Gln Phe Leu Ile Thr Ala Cys Gly Thr Gl - #u Val Ala Leu Asp Leu           #               415                                                           - CTG GAA GAG CAG GTT ATC TCT GAG GTG GGG GT - #C ATG ACC TGG GTG AGC         1416                                                                          Leu Glu Glu Gln Val Ile Ser Glu Val Gly Va - #l Met Thr Trp Val Ser           #           430                                                               - CGA CAG AAG CAG GAG ATG GTG GAG AGC AAC TC - #C AAA ATC ATC ATC CTG         1464                                                                          Arg Gln Lys Gln Glu Met Val Glu Ser Asn Se - #r Lys Ile Ile Ile Leu           #       445                                                                   - TGT TCC CGA GGC ACC CAA GCA AAG TGG AAA GC - #T ATC TTG GGT TGG GCT         1512                                                                          Cys Ser Arg Gly Thr Gln Ala Lys Trp Lys Al - #a Ile Leu Gly Trp Ala           #   460                                                                       - GAG CCT GCT GTC CAG CTA CGG TGT GAC CAC TG - #G AAG CCT GCT GGG GAC         1560                                                                          Glu Pro Ala Val Gln Leu Arg Cys Asp His Tr - #p Lys Pro Ala Gly Asp           465                 4 - #70                 4 - #75                 4 -       #80                                                                           - CTT TTC ACT GCA GCC ATG AAC ATG ATC CTG CC - #A GAC TTC AAG AGG CCA         1608                                                                          Leu Phe Thr Ala Ala Met Asn Met Ile Leu Pr - #o Asp Phe Lys Arg Pro           #               495                                                           - GCC TGC TTC GGC ACC TAC GTT GTT TGC TAC TT - #C AGT GGC ATC TGT AGT         1656                                                                          Ala Cys Phe Gly Thr Tyr Val Val Cys Tyr Ph - #e Ser Gly Ile Cys Ser           #           510                                                               - GAG AGG GAT GTC CCC GAC CTC TTC AAC ATC AC - #C TCC AGG TAC CCA CTC         1704                                                                          Glu Arg Asp Val Pro Asp Leu Phe Asn Ile Th - #r Ser Arg Tyr Pro Leu           #       525                                                                   - ATG GAC AGA TTT GAG GAG GTT TAC TTC CGG AT - #C CAG GAC CTG GAG ATG         1752                                                                          Met Asp Arg Phe Glu Glu Val Tyr Phe Arg Il - #e Gln Asp Leu Glu Met           #   540                                                                       - TTT GAA CCC GGC CGG ATG CAC CAT GTC AGA GA - #G CTC ACA GGG GAC AAT         1800                                                                          Phe Glu Pro Gly Arg Met His His Val Arg Gl - #u Leu Thr Gly Asp Asn           545                 5 - #50                 5 - #55                 5 -       #60                                                                           - TAC CTG CAG AGC CCT AGT GGC CGG CAG CTC AA - #G GAG GCT GTG CTT AGG         1848                                                                          Tyr Leu Gln Ser Pro Ser Gly Arg Gln Leu Ly - #s Glu Ala Val Leu Arg           #               575                                                           - TTC CAG GAG TGG CAA ACC CAG TGC CCC GAC TG - #G TTC GAG CGT GAG AAC         1896                                                                          Phe Gln Glu Trp Gln Thr Gln Cys Pro Asp Tr - #p Phe Glu Arg Glu Asn           #           590                                                               - CTC TGC TTA GCT GAT GGC CAA GAT CTT CCC TC - #C CTG GAT GAA GAA GTG         1944                                                                          Leu Cys Leu Ala Asp Gly Gln Asp Leu Pro Se - #r Leu Asp Glu Glu Val           #       605                                                                   - TTT GAA GAC CCA CTG CTG CCA CCA GGG GGA GG - #A ATT GTC AAA CAG CAG         1992                                                                          Phe Glu Asp Pro Leu Leu Pro Pro Gly Gly Gl - #y Ile Val Lys Gln Gln           #   620                                                                       - CCC CTG GTG CGG GAA CTC CCA TCT GAC GGC TG - #C CTT GTG GTA GAT GTC         2040                                                                          Pro Leu Val Arg Glu Leu Pro Ser Asp Gly Cy - #s Leu Val Val Asp Val           625                 6 - #30                 6 - #35                 6 -       #40                                                                           - TGT GTC AGT GAG GAA GAA AGT AGA ATG GCA AA - #G CTG GAC CCT CAG CTA         2088                                                                          Cys Val Ser Glu Glu Glu Ser Arg Met Ala Ly - #s Leu Asp Pro Gln Leu           #               655                                                           - TGG CCA CAG AGA GAG CTA GTG GCT CAC ACC CT - #C CAA AGC ATG GTG CTG         2136                                                                          Trp Pro Gln Arg Glu Leu Val Ala His Thr Le - #u Gln Ser Met Val Leu           #           670                                                               - CCA GCA GAG CAG GTC CCT GCA GCT CAT GTG GT - #G GAG CCT CTC CAT CTC         2184                                                                          Pro Ala Glu Gln Val Pro Ala Ala His Val Va - #l Glu Pro Leu His Leu           #       685                                                                   - CCA GAC GGC AGT GGA GCA GCT GCC CAG CTG CC - #C ATG ACA GAG GAC AGC         2232                                                                          Pro Asp Gly Ser Gly Ala Ala Ala Gln Leu Pr - #o Met Thr Glu Asp Ser           #   700                                                                       - GAG GCT TGC CCG CTG CTG GGG GTC CAG AGG AA - #C AGC ATC CTT TGC CTC         2280                                                                          Glu Ala Cys Pro Leu Leu Gly Val Gln Arg As - #n Ser Ile Leu Cys Leu           705                 7 - #10                 7 - #15                 7 -       #20                                                                           - CCC GTG GAC TCA GAT GAC TTG CCA CTC TGT AG - #C ACC CCA ATG ATG TCA         2328                                                                          Pro Val Asp Ser Asp Asp Leu Pro Leu Cys Se - #r Thr Pro Met Met Ser           #               735                                                           - CCT GAC CAC CTC CAA GGC GAT GCA AGA GAG CA - #G CTA GAA AGC CTA ATG         2376                                                                          Pro Asp His Leu Gln Gly Asp Ala Arg Glu Gl - #n Leu Glu Ser Leu Met           #           750                                                               - CTC TCG GTG CTG CAG CAG AGC CTG AGT GGA CA - #G CCC CTG GAG AGC TGG         2424                                                                          Leu Ser Val Leu Gln Gln Ser Leu Ser Gly Gl - #n Pro Leu Glu Ser Trp           #       765                                                                   - CCG AGG CCA GAG GTG GTC CTC GAG GGC TGC AC - #A CCC TCT GAG GAG GAG         2472                                                                          Pro Arg Pro Glu Val Val Leu Glu Gly Cys Th - #r Pro Ser Glu Glu Glu           #   780                                                                       - CAG CGG CAG TCG GTG CAG TCG GAC CAG GGC TA - #C ATC TCC AGG AGC TCC         2520                                                                          Gln Arg Gln Ser Val Gln Ser Asp Gln Gly Ty - #r Ile Ser Arg Ser Ser           785                 7 - #90                 7 - #95                 8 -       #00                                                                           - CCG CAG CCC CCC GAG TGG CTC ACG GAG GAG GA - #A GAG CTA GAA CTG GGT         2568                                                                          Pro Gln Pro Pro Glu Trp Leu Thr Glu Glu Gl - #u Glu Leu Glu Leu Gly           #               815                                                           - GAG CCC GTT GAG TCT CTC TCT CCT GAG GAA CT - #A CGG AGC CTG AGG AAG         2616                                                                          Glu Pro Val Glu Ser Leu Ser Pro Glu Glu Le - #u Arg Ser Leu Arg Lys           #           830                                                               - CTC CAG AGG CAG CTT TTC TTC TGG GAG CTC GA - #G AAG AAC CCT GGC TGG         2664                                                                          Leu Gln Arg Gln Leu Phe Phe Trp Glu Leu Gl - #u Lys Asn Pro Gly Trp           #       845                                                                   - AAC AGC TTG GAG CCA CGG AGA CCC ACC CCA GA - #A GAG CAG AAT CCC TCC         2712                                                                          Asn Ser Leu Glu Pro Arg Arg Pro Thr Pro Gl - #u Glu Gln Asn Pro Ser           #   860                                                                       - TAG GCCTCCTGAG CCTGCTACTT AAGAGGGTGT ATATTGTACT CTGTGTGTG - #C              2765                                                                           *                                                                            865                                                                           - GTGCGTGTGT GTGTGTGTGT GTGTGTGTGT GTGCGTGTGT GTGTGTGTGT GT - #GTGTGTGT       2825                                                                          - GTGTGTGTAG TGCCCGGCTT AGAAATGTGA ACATCTGAAT CTGACATAGT GT - #TGTATACC       2885                                                                          - TGAAGTCCCA GCACTTGGGA ACTGAGACTT GATGATCTCC TGAAGCCAGG TG - #TTCAGGGC       2945                                                                          - CAGTGTGAAA ACATAGCAAG ACCTCAGAGA AATCAATGCA GACATCTTGG TA - #CTGATCCC       3005                                                                          - TAAACACACC CCTTTCCCTG ATAACCCGAC ATGAGCATCT GGTCATCATT GC - #ACAAGAAT       3065                                                                          - CCACAGCCCG TTCCCAGAGC TCATAGCCAA GTGTGTTGCT CATTCCTTGA AT - #ATTTATTC       3125                                                                          - TGTACCTACT ATTCATCAGA CATTTGGAAT TCAAAAACAA GTTACATGAC AC - #AGCCTTAG       3185                                                                          - CCACTAAGAA GCTTAAAATT CGGTAAGGAT GTAAAATTAG CCAGGATGAA TA - #GAGGGCTG       3245                                                                          #                 328 - #8GGTCGTCT CGTTCCAGTC GAC                             - (2) INFORMATION FOR SEQ ID NO:2:                                            -      (i) SEQUENCE CHARACTERISTICS:                                                    (A) LENGTH:  864 ami - #no acids                                              (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                 - Met Ala Ile Arg Arg Cys Trp Pro Arg Val Va - #l Pro Gly Pro Ala Leu         #                 15                                                          - Gly Trp Leu Leu Leu Leu Leu Asn Val Leu Al - #a Pro Gly Arg Ala Ser         #             30                                                              - Pro Arg Leu Leu Asp Phe Pro Ala Pro Val Cy - #s Ala Gln Glu Gly Leu         #         45                                                                  - Ser Cys Arg Val Lys Asn Ser Thr Cys Leu As - #p Asp Ser Trp Ile His         #     60                                                                      - Pro Lys Asn Leu Thr Pro Ser Ser Pro Lys As - #n Ile Tyr Ile Asn Leu         # 80                                                                          - Ser Val Ser Ser Thr Gln His Gly Glu Leu Va - #l Pro Val Leu His Val         #                 95                                                          - Glu Trp Thr Leu Gln Thr Asp Ala Ser Ile Le - #u Tyr Leu Glu Gly Ala         #           110                                                               - Glu Leu Ser Val Leu Gln Leu Asn Thr Asn Gl - #u Arg Leu Cys Val Lys         #       125                                                                   - Phe Gln Phe Leu Ser Met Leu Gln His His Ar - #g Lys Arg Trp Arg Phe         #   140                                                                       - Ser Phe Ser His Phe Val Val Asp Pro Gly Gl - #n Glu Tyr Glu Val Thr         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Val His His Leu Pro Lys Pro Ile Pro Asp Gl - #y Asp Pro Asn His Lys         #               175                                                           - Ser Lys Ile Ile Phe Val Pro Asp Cys Glu As - #p Ser Lys Met Lys Met         #           190                                                               - Thr Thr Ser Cys Val Ser Ser Gly Ser Leu Tr - #p Asp Pro Asn Ile Thr         #       205                                                                   - Val Glu Thr Leu Asp Thr Gln His Leu Arg Va - #l Asp Phe Thr Leu Trp         #   220                                                                       - Asn Glu Ser Thr Pro Tyr Gln Val Leu Leu Gl - #u Ser Phe Ser Asp Ser         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Glu Asn His Ser Cys Phe Asp Val Val Lys Gl - #n Ile Phe Ala Pro Arg         #               255                                                           - Gln Glu Glu Phe His Gln Arg Ala Asn Val Th - #r Phe Thr Leu Ser Lys         #           270                                                               - Phe His Trp Cys Cys His His His Val Gln Va - #l Gln Pro Phe Phe Ser         #       285                                                                   - Ser Cys Leu Asn Asp Cys Leu Arg His Ala Va - #l Thr Val Pro Cys Pro         #   300                                                                       - Val Ile Ser Asn Thr Thr Val Pro Lys Pro Va - #l Ala Asp Tyr Ile Pro         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Leu Trp Val Tyr Gly Leu Ile Thr Leu Ile Al - #a Ile Leu Leu Val Gly         #               335                                                           - Ser Val Ile Val Leu Ile Ile Cys Met Thr Tr - #p Arg Leu Ser Gly Ala         #           350                                                               - Asp Gln Glu Lys His Gly Asp Asp Ser Lys Il - #e Asn Gly Ile Leu Pro         #       365                                                                   - Val Ala Asp Leu Thr Pro Pro Pro Leu Arg Pr - #o Arg Lys Val Trp Ile         #   380                                                                       - Val Tyr Ser Ala Asp His Pro Leu Tyr Val Gl - #u Val Val Leu Lys Phe         385                 3 - #90                 3 - #95                 4 -       #00                                                                           - Ala Gln Phe Leu Ile Thr Ala Cys Gly Thr Gl - #u Val Ala Leu Asp Leu         #               415                                                           - Leu Glu Glu Gln Val Ile Ser Glu Val Gly Va - #l Met Thr Trp Val Ser         #           430                                                               - Arg Gln Lys Gln Glu Met Val Glu Ser Asn Se - #r Lys Ile Ile Ile Leu         #       445                                                                   - Cys Ser Arg Gly Thr Gln Ala Lys Trp Lys Al - #a Ile Leu Gly Trp Ala         #   460                                                                       - Glu Pro Ala Val Gln Leu Arg Cys Asp His Tr - #p Lys Pro Ala Gly Asp         465                 4 - #70                 4 - #75                 4 -       #80                                                                           - Leu Phe Thr Ala Ala Met Asn Met Ile Leu Pr - #o Asp Phe Lys Arg Pro         #               495                                                           - Ala Cys Phe Gly Thr Tyr Val Val Cys Tyr Ph - #e Ser Gly Ile Cys Ser         #           510                                                               - Glu Arg Asp Val Pro Asp Leu Phe Asn Ile Th - #r Ser Arg Tyr Pro Leu         #       525                                                                   - Met Asp Arg Phe Glu Glu Val Tyr Phe Arg Il - #e Gln Asp Leu Glu Met         #   540                                                                       - Phe Glu Pro Gly Arg Met His His Val Arg Gl - #u Leu Thr Gly Asp Asn         545                 5 - #50                 5 - #55                 5 -       #60                                                                           - Tyr Leu Gln Ser Pro Ser Gly Arg Gln Leu Ly - #s Glu Ala Val Leu Arg         #               575                                                           - Phe Gln Glu Trp Gln Thr Gln Cys Pro Asp Tr - #p Phe Glu Arg Glu Asn         #           590                                                               - Leu Cys Leu Ala Asp Gly Gln Asp Leu Pro Se - #r Leu Asp Glu Glu Val         #       605                                                                   - Phe Glu Asp Pro Leu Leu Pro Pro Gly Gly Gl - #y Ile Val Lys Gln Gln         #   620                                                                       - Pro Leu Val Arg Glu Leu Pro Ser Asp Gly Cy - #s Leu Val Val Asp Val         625                 6 - #30                 6 - #35                 6 -       #40                                                                           - Cys Val Ser Glu Glu Glu Ser Arg Met Ala Ly - #s Leu Asp Pro Gln Leu         #               655                                                           - Trp Pro Gln Arg Glu Leu Val Ala His Thr Le - #u Gln Ser Met Val Leu         #           670                                                               - Pro Ala Glu Gln Val Pro Ala Ala His Val Va - #l Glu Pro Leu His Leu         #       685                                                                   - Pro Asp Gly Ser Gly Ala Ala Ala Gln Leu Pr - #o Met Thr Glu Asp Ser         #   700                                                                       - Glu Ala Cys Pro Leu Leu Gly Val Gln Arg As - #n Ser Ile Leu Cys Leu         705                 7 - #10                 7 - #15                 7 -       #20                                                                           - Pro Val Asp Ser Asp Asp Leu Pro Leu Cys Se - #r Thr Pro Met Met Ser         #               735                                                           - Pro Asp His Leu Gln Gly Asp Ala Arg Glu Gl - #n Leu Glu Ser Leu Met         #           750                                                               - Leu Ser Val Leu Gln Gln Ser Leu Ser Gly Gl - #n Pro Leu Glu Ser Trp         #       765                                                                   - Pro Arg Pro Glu Val Val Leu Glu Gly Cys Th - #r Pro Ser Glu Glu Glu         #   780                                                                       - Gln Arg Gln Ser Val Gln Ser Asp Gln Gly Ty - #r Ile Ser Arg Ser Ser         785                 7 - #90                 7 - #95                 8 -       #00                                                                           - Pro Gln Pro Pro Glu Trp Leu Thr Glu Glu Gl - #u Glu Leu Glu Leu Gly         #               815                                                           - Glu Pro Val Glu Ser Leu Ser Pro Glu Glu Le - #u Arg Ser Leu Arg Lys         #           830                                                               - Leu Gln Arg Gln Leu Phe Phe Trp Glu Leu Gl - #u Lys Asn Pro Gly Trp         #       845                                                                   - Asn Ser Leu Glu Pro Arg Arg Pro Thr Pro Gl - #u Glu Gln Asn Pro Ser         #   860                                                                       -  *                                                                          865                                                                           - (2) INFORMATION FOR SEQ ID NO:3:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 8 amino                                                           (B) TYPE: amino acid                                                          (C) STRANDEDNESS: Not R - #elevant                                            (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (vii) IMMEDIATE SOURCE:                                                            (B) CLONE: FLAG.sub.-- - # peptide                                  -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                 - Asp Tyr Lys Asp Asp Asp Asp Lys                                               1               5                                                           - (2) INFORMATION FOR SEQ ID NO:4:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 212 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: Not R - #elevant                                            (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Human                                                 -    (vii) IMMEDIATE SOURCE:                                                            (B) CLONE: IgG1 Fc                                                  -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                 - Arg Ser Cys Asp Lys Thr His Thr Cys Pro Pr - #o Cys Pro Ala Pro Glu         #                15                                                           - Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pr - #o Pro Lys Pro Lys Asp         #            30                                                               - Thr Leu Met Ile Ser Arg Thr Pro Glu Val Th - #r Cys Val Val Val Asp         #        45                                                                   - Val Ser His Glu Asp Pro Glu Val Lys Phe As - #n Trp Tyr Val Asp Gly         #    60                                                                       - Val Glu Val His Asn Ala Lys Thr Lys Pro Ar - #g Glu Glu Gln Tyr Asn         #80                                                                           - Ser Thr Tyr Arg Val Val Ser Val Leu Thr Va - #l Leu His Gln Asp Trp         #                95                                                           - Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Se - #r Asn Lys Ala Leu Pro         #           110                                                               - Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Ly - #s Gly Gln Pro Arg Glu         #       125                                                                   - Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg As - #p Glu Leu Thr Lys Asn         #   140                                                                       - Gln Val Ser Leu Thr Cys Leu Val Lys Gly Ph - #e Tyr Pro Ser Asp Ile         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Gl - #u Asn Asn Tyr Lys Thr         #               175                                                           - Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Ph - #e Phe Leu Tyr Ser Lys         #           190                                                               - Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gl - #y Asn Val Phe Ser Cys         #       205                                                                   - Ser Val Met His                                                                 210                                                                       - (2) INFORMATION FOR SEQ ID NO:5:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 14 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: Not R - #elevant                                            (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (vii) IMMEDIATE SOURCE:                                                            (B) CLONE: Polylinker                                               -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                 - Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gl - #y Gly Gly Gly                 #                 10                                                          - (2) INFORMATION FOR SEQ ID NO:6:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 498 base                                                          (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA to mRNA                                        -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Murine CT - #LA-8                                     -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 14..490                                               -     (ix) FEATURE:                                                                     (A) NAME/KEY: sig.sub.-- - #peptide                                           (B) LOCATION: 14..88                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: mat.sub.-- - #peptide                                           (B) LOCATION: 89..487                                               -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                 - GTCGACCCCC ACC ATG TTC CAT GTT TCT TTT AGA TA - #T ATC TTT GGA ATT            49                                                                          #His Val Ser Phe Arg Tyr Ile Phe Gly Ile                                      15                                                                            - CCT CCA CTG ATC CTT GTT CTG CTG CCT GTC AC - #T AGT TCT GCG GTA CTC           97                                                                          Pro Pro Leu Ile Leu Val Leu Leu Pro Val Th - #r Ser Ser Ala Val Leu           #             1                                                               - ATC CCT CAA AGT TCA GCG TGT CCA AAC ACT GA - #G GCC AAG GAC TTC CTC          145                                                                          Ile Pro Gln Ser Ser Ala Cys Pro Asn Thr Gl - #u Ala Lys Asp Phe Leu           #      15                                                                     - CAG AAT GTG AAG GTC AAC CTC AAA GTC TTT AA - #C TCC CTT GGC GCA AAA          193                                                                          Gln Asn Val Lys Val Asn Leu Lys Val Phe As - #n Ser Leu Gly Ala Lys           # 35                                                                          - GTG AGC TCC AGA AGG CCC TCA GAC TAC CTC AA - #C CGT TCC ACG TCA CCC          241                                                                          Val Ser Ser Arg Arg Pro Ser Asp Tyr Leu As - #n Arg Ser Thr Ser Pro           #                 50                                                          - TGG ACT CTC CAC CGC AAT GAA GAC CCT GAT AG - #A TAT CCC TCT GTG ATC          289                                                                          Trp Thr Leu His Arg Asn Glu Asp Pro Asp Ar - #g Tyr Pro Ser Val Ile           #             65                                                              - TGG GAA GCT CAG TGC CGC CAC CAG CGC TGT GT - #C AAT GCG GAG GGA AAG          337                                                                          Trp Glu Ala Gln Cys Arg His Gln Arg Cys Va - #l Asn Ala Glu Gly Lys           #         80                                                                  - CTG GAC CAC CAC ATG AAT TCT GTT CTC ATC CA - #G CAA GAG ATC CTG GTC          385                                                                          Leu Asp His His Met Asn Ser Val Leu Ile Gl - #n Gln Glu Ile Leu Val           #     95                                                                      - CTG AAG AGG GAG CCT GAG AGC TGC CCC TTC AC - #T TTC AGG GTC GAG AAG          433                                                                          Leu Lys Arg Glu Pro Glu Ser Cys Pro Phe Th - #r Phe Arg Val Glu Lys           100                 1 - #05                 1 - #10                 1 -       #15                                                                           - ATG CTG GTG GGT GTG GGC TGC ACC TGC GTG GC - #C TCG ATT GTC CGC CAT          481                                                                          Met Leu Val Gly Val Gly Cys Thr Cys Val Al - #a Ser Ile Val Arg His           #               130                                                           #  498             CGC                                                        Ala Ser  *                                                                    - (2) INFORMATION FOR SEQ ID NO:7:                                            -      (i) SEQUENCE CHARACTERISTICS:                                                    (A) LENGTH:  158 ami - #no acids                                              (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                 - Met Phe His Val Ser Phe Arg Tyr Ile Phe Gl - #y Ile Pro Pro Leu Ile         - #10                                                                         - Leu Val Leu Leu Pro Val Thr Ser Ser Ala Va - #l Leu Ile Pro Gln Ser         #1               5                                                            - Ser Ala Cys Pro Asn Thr Glu Ala Lys Asp Ph - #e Leu Gln Asn Val Lys         #         20                                                                  - Val Asn Leu Lys Val Phe Asn Ser Leu Gly Al - #a Lys Val Ser Ser Arg         #     35                                                                      - Arg Pro Ser Asp Tyr Leu Asn Arg Ser Thr Se - #r Pro Trp Thr Leu His         # 55                                                                          - Arg Asn Glu Asp Pro Asp Arg Tyr Pro Ser Va - #l Ile Trp Glu Ala Gln         #                 70                                                          - Cys Arg His Gln Arg Cys Val Asn Ala Glu Gl - #y Lys Leu Asp His His         #             85                                                              - Met Asn Ser Val Leu Ile Gln Gln Glu Ile Le - #u Val Leu Lys Arg Glu         #        100                                                                  - Pro Glu Ser Cys Pro Phe Thr Phe Arg Val Gl - #u Lys Met Leu Val Gly         #   115                                                                       - Val Gly Cys Thr Cys Val Ala Ser Ile Val Ar - #g His Ala Ser                 120                 1 - #25                 1 - #30                           - (2) INFORMATION FOR SEQ ID NO:8:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 151 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: Not R - #elevant                                            (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -     (vi) ORIGINAL SOURCE:                                                   #Saimiri  (A) ORGANISM: Herpesvirus                                                     (B) STRAIN: ORF13                                                   -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                 - Met Thr Phe Arg Met Thr Ser Leu Val Leu Le - #u Leu Leu Leu Ser Ile         #                15                                                           - Asp Cys Ile Val Lys Ser Glu Ile Thr Ser Al - #a Gln Thr Pro Arg Cys         #            30                                                               - Leu Ala Ala Asn Asn Ser Phe Pro Arg Ser Va - #l Met Val Thr Leu Ser         #        45                                                                   - Ile Arg Asn Trp Asn Thr Ser Ser Lys Arg Al - #a Ser Asp Tyr Tyr Asn         #    60                                                                       - Arg Ser Thr Ser Pro Trp Thr Leu His Arg As - #n Glu Asp Gln Asp Arg         #80                                                                           - Tyr Pro Ser Val Ile Trp Glu Ala Lys Cys Ar - #g Tyr Leu Gly Cys Val         #                95                                                           - Asn Ala Asp Gly Asn Val Asp Tyr His Met As - #n Ser Val Pro Ile Gln         #           110                                                               - Gln Glu Ile Leu Val Val Arg Lys Gly His Gl - #n Pro Cys Pro Asn Ser         #       125                                                                   - Phe Arg Leu Glu Lys Met Leu Val Thr Val Gl - #y Cys Thr Cys Val Thr         #   140                                                                       - Pro Ile Val His Asn Val Asp                                                 145                 1 - #50                                                   - (2) INFORMATION FOR SEQ ID NO:9:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #pairs    (A) LENGTH: 3223 base                                                         (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: cDNA to mRNA                                        -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Human                                                 #(hCTLA8 receptor)IN: IL-17 R                                                 -     (ix) FEATURE:                                                                     (A) NAME/KEY: CDS                                                             (B) LOCATION: 93..2693                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                 - GGGAGACCGG AATTCCGGGA AAAGAAAGCC TCAGAACGTT CGCTCGCTGC GT - #CCCCAGCC         60                                                                          - GGGGCCGAGC CCTCCGCGAC GCCACCCGGG CC ATG GGG GCC GCA - # CGC AGC CCG          113                                                                          #Met Gly Ala Ala Arg Ser Pro                                                  #  1               5                                                          - CCG TCC GCT GTC CCG GGG CCC CTG CTG GGG CT - #G CTC CTG CTG CTC CTG          161                                                                          Pro Ser Ala Val Pro Gly Pro Leu Leu Gly Le - #u Leu Leu Leu Leu Leu           #         20                                                                  - GGC GTG CTG GCC CCG GGT GGC GCC TCC CTG CG - #A CTC CTG GAC CAC CGG          209                                                                          Gly Val Leu Ala Pro Gly Gly Ala Ser Leu Ar - #g Leu Leu Asp His Arg           #     35                                                                      - GCG CTG GTC TGC TCC CAG CCG GGG CTA AAC TG - #C ACG GTC AAG AAT AGT          257                                                                          Ala Leu Val Cys Ser Gln Pro Gly Leu Asn Cy - #s Thr Val Lys Asn Ser           # 55                                                                          - ACC TGC CTG GAT GAC AGC TGG ATT CAC CCT CG - #A AAC CTG ACC CCC TCC          305                                                                          Thr Cys Leu Asp Asp Ser Trp Ile His Pro Ar - #g Asn Leu Thr Pro Ser           #                 70                                                          - TCC CCA AAG GAC CTG CAG ATC CAG CTG CAC TT - #T GCC CAC ACC CAA CAA          353                                                                          Ser Pro Lys Asp Leu Gln Ile Gln Leu His Ph - #e Ala His Thr Gln Gln           #             85                                                              - GGA GAC CTG TTC CCC GTG GCT CAC ATC GAA TG - #G ACA CTG CAG ACA GAC          401                                                                          Gly Asp Leu Phe Pro Val Ala His Ile Glu Tr - #p Thr Leu Gln Thr Asp           #        100                                                                  - GCC AGC ATC CTG TAC CTC GAG GGT GCA GAG TT - #A TCT GTC CTG CAG CTG          449                                                                          Ala Ser Ile Leu Tyr Leu Glu Gly Ala Glu Le - #u Ser Val Leu Gln Leu           #   115                                                                       - AAC ACC AAT GAA CGT TTG TGC GTC AGG TTT GA - #G TTT CTG TCC AAA CTG          497                                                                          Asn Thr Asn Glu Arg Leu Cys Val Arg Phe Gl - #u Phe Leu Ser Lys Leu           120                 1 - #25                 1 - #30                 1 -       #35                                                                           - AGG CAT CAC CAC AGG CGG TGG CGT TTT ACC TT - #C AGC CAC TTT GTG GTT          545                                                                          Arg His His His Arg Arg Trp Arg Phe Thr Ph - #e Ser His Phe Val Val           #               150                                                           - GAC CCT GAC CAG GAA TAT GAG GTG ACC GTT CA - #C CAC CTG CCC AAG CCC          593                                                                          Asp Pro Asp Gln Glu Tyr Glu Val Thr Val Hi - #s His Leu Pro Lys Pro           #           165                                                               - ATC CCT GAT GGG GAC CCA AAC CAC CAG TCC AA - #G AAT TTC CTT GTG CCT          641                                                                          Ile Pro Asp Gly Asp Pro Asn His Gln Ser Ly - #s Asn Phe Leu Val Pro           #       180                                                                   - GAC TGT GAG CAC GCC AGG ATG AAG GTA ACC AC - #G CCA TGC ATG AGC TCA          689                                                                          Asp Cys Glu His Ala Arg Met Lys Val Thr Th - #r Pro Cys Met Ser Ser           #   195                                                                       - GGC AGC CTG TGG GAC CCC AAC ATC ACC GTG GA - #G ACC CTG GAG GCC CAC          737                                                                          Gly Ser Leu Trp Asp Pro Asn Ile Thr Val Gl - #u Thr Leu Glu Ala His           200                 2 - #05                 2 - #10                 2 -       #15                                                                           - CAG CTG CGT GTG AGC TTC ACC CTG TGG AAC GA - #A TCT ACC CAT TAC CAG          785                                                                          Gln Leu Arg Val Ser Phe Thr Leu Trp Asn Gl - #u Ser Thr His Tyr Gln           #               230                                                           - ATC CTG CTG ACC AGT TTT CCG CAC ATG GAG AA - #C CAC AGT TGC TTT GAG          833                                                                          Ile Leu Leu Thr Ser Phe Pro His Met Glu As - #n His Ser Cys Phe Glu           #           245                                                               - CAC ATG CAC CAC ATA CCT GCG CCC AGA CCA GA - #A GAG TTC CAC CAG CGA          881                                                                          His Met His His Ile Pro Ala Pro Arg Pro Gl - #u Glu Phe His Gln Arg           #       260                                                                   - TCC AAC GTC ACA CTC ACT CTA CGC AAC CTT AA - #A GGG TGC TGT CGC CAC          929                                                                          Ser Asn Val Thr Leu Thr Leu Arg Asn Leu Ly - #s Gly Cys Cys Arg His           #   275                                                                       - CAA GTG CAG ATC CAG CCC TTC TTC AGC AGC TG - #C CTC AAT GAC TGC CTC          977                                                                          Gln Val Gln Ile Gln Pro Phe Phe Ser Ser Cy - #s Leu Asn Asp Cys Leu           280                 2 - #85                 2 - #90                 2 -       #95                                                                           - AGA CAC TCC GCG ACT GTT TCC TGC CCA GAA AT - #G CCA GAC ACT CCA GAA         1025                                                                          Arg His Ser Ala Thr Val Ser Cys Pro Glu Me - #t Pro Asp Thr Pro Glu           #               310                                                           - CCA ATT CCG GAC TAC ATG CCC CTG TGG GTG TA - #C TGG TTC ATC ACG GGC         1073                                                                          Pro Ile Pro Asp Tyr Met Pro Leu Trp Val Ty - #r Trp Phe Ile Thr Gly           #           325                                                               - ATC TCC ATC CTG CTG GTG GGC TCC GTC ATC CT - #G CTC ATC GTC TGC ATG         1121                                                                          Ile Ser Ile Leu Leu Val Gly Ser Val Ile Le - #u Leu Ile Val Cys Met           #       340                                                                   - ACC TGG AGG CTA GCT GGG CCT GGA AGT GAA AA - #A TAC AGT GAT GAC ACC         1169                                                                          Thr Trp Arg Leu Ala Gly Pro Gly Ser Glu Ly - #s Tyr Ser Asp Asp Thr           #   355                                                                       - AAA TAC ACC GAT GGC CTG CCT GCG GCT GAC CT - #G ATC CCC CCA CCG CTG         1217                                                                          Lys Tyr Thr Asp Gly Leu Pro Ala Ala Asp Le - #u Ile Pro Pro Pro Leu           360                 3 - #65                 3 - #70                 3 -       #75                                                                           - AAG CCC AGG AAG GTC TGG ATC ATC TAC TCA GC - #C GAC CAC CCC CTC TAC         1265                                                                          Lys Pro Arg Lys Val Trp Ile Ile Tyr Ser Al - #a Asp His Pro Leu Tyr           #               390                                                           - GTG GAC GTG GTC CTG AAA TTC GCC CAG TTC CT - #G CTC ACC GCC TGC GGC         1313                                                                          Val Asp Val Val Leu Lys Phe Ala Gln Phe Le - #u Leu Thr Ala Cys Gly           #           405                                                               - ACG GAA GTG GCC CTG GAC CTG CTG GAA GAG CA - #G GCC ATC TCG GAG GCA         1361                                                                          Thr Glu Val Ala Leu Asp Leu Leu Glu Glu Gl - #n Ala Ile Ser Glu Ala           #       420                                                                   - GGA GTC ATG ACC TGG GTG GGC CGT CAG AAG CA - #G GAG ATG GTG GAG AGC         1409                                                                          Gly Val Met Thr Trp Val Gly Arg Gln Lys Gl - #n Glu Met Val Glu Ser           #   435                                                                       - AAC TCT AAG ATC ATC GTC CTG TGC TCC CGC GG - #C ACG CGC GCC AAG TGG         1457                                                                          Asn Ser Lys Ile Ile Val Leu Cys Ser Arg Gl - #y Thr Arg Ala Lys Trp           440                 4 - #45                 4 - #50                 4 -       #55                                                                           - CAG GCG CTC CTG GGC CGG GGG GCG CCT GTG CG - #G CTG CGC TGC GAC CAC         1505                                                                          Gln Ala Leu Leu Gly Arg Gly Ala Pro Val Ar - #g Leu Arg Cys Asp His           #               470                                                           - GGA AAG CCC GTG GGG GAC CTG TTC ACT GCA GC - #C ATG AAC ATG ATC CTC         1553                                                                          Gly Lys Pro Val Gly Asp Leu Phe Thr Ala Al - #a Met Asn Met Ile Leu           #           485                                                               - CCG GAC TTC AAG AGG CCA GCC TGC TTC GGC AC - #C TAC GTA GTC TGC TAC         1601                                                                          Pro Asp Phe Lys Arg Pro Ala Cys Phe Gly Th - #r Tyr Val Val Cys Tyr           #       500                                                                   - TTC AGC GAG GTC AGC TGT GAC GGC GAC GTC CC - #C GAC CTG TTC GGC GCG         1649                                                                          Phe Ser Glu Val Ser Cys Asp Gly Asp Val Pr - #o Asp Leu Phe Gly Ala           #   515                                                                       - GCG CCG CGG TAC CCG CTC ATG GAC AGG TTC GA - #G GAG GTG TAC TTC CGC         1697                                                                          Ala Pro Arg Tyr Pro Leu Met Asp Arg Phe Gl - #u Glu Val Tyr Phe Arg           520                 5 - #25                 5 - #30                 5 -       #35                                                                           - ATC CAG GAC CTG GAG ATG TTC CAG CCG GGC CG - #C ATG CAC CGC GTA GGG         1745                                                                          Ile Gln Asp Leu Glu Met Phe Gln Pro Gly Ar - #g Met His Arg Val Gly           #               550                                                           - GAG CTG TCG GGG GAC AAC TAC CTG CGG AGC CC - #G GGC GGC AGG CAG CTC         1793                                                                          Glu Leu Ser Gly Asp Asn Tyr Leu Arg Ser Pr - #o Gly Gly Arg Gln Leu           #           565                                                               - CGC GCC GCC CTG GAC AGG TTC CGG GAC TGG CA - #G GTC CGC TGT CCC GAC         1841                                                                          Arg Ala Ala Leu Asp Arg Phe Arg Asp Trp Gl - #n Val Arg Cys Pro Asp           #       580                                                                   - TGG TTC GAA TGT GAG AAC CTC TAC TCA GCA GA - #T GAC CAG GAT GCC CCG         1889                                                                          Trp Phe Glu Cys Glu Asn Leu Tyr Ser Ala As - #p Asp Gln Asp Ala Pro           #   595                                                                       - TCC CTG GAC GAA GAG GTG TTT GAG GAG CCA CT - #G CTG CCT CCG GGA ACC         1937                                                                          Ser Leu Asp Glu Glu Val Phe Glu Glu Pro Le - #u Leu Pro Pro Gly Thr           600                 6 - #05                 6 - #10                 6 -       #15                                                                           - GGC ATC GTG AAG CGG GCG CCC CTG GTG CGC GA - #G CCT GGC TCC CAG GCC         1985                                                                          Gly Ile Val Lys Arg Ala Pro Leu Val Arg Gl - #u Pro Gly Ser Gln Ala           #               630                                                           - TGC CTG GCC ATA GAC CCG CTG GTC GGG GAG GA - #A GGA GGA GCA GCA GTG         2033                                                                          Cys Leu Ala Ile Asp Pro Leu Val Gly Glu Gl - #u Gly Gly Ala Ala Val           #           645                                                               - GCA AAG CTG GAA CCT CAC CTG CAG CCC CGG GG - #T CAG CCA GCG CCG CAG         2081                                                                          Ala Lys Leu Glu Pro His Leu Gln Pro Arg Gl - #y Gln Pro Ala Pro Gln           #       660                                                                   - CCC CTC CAC ACC CTG GTG CTC GCC GCA GAG GA - #G GGG GCC CTG GTG GCC         2129                                                                          Pro Leu His Thr Leu Val Leu Ala Ala Glu Gl - #u Gly Ala Leu Val Ala           #   675                                                                       - GCG GTG GAG CCT GGG CCC CTG GCT GAC GGT GC - #C GCA GTC CGG CTG GCA         2177                                                                          Ala Val Glu Pro Gly Pro Leu Ala Asp Gly Al - #a Ala Val Arg Leu Ala           680                 6 - #85                 6 - #90                 6 -       #95                                                                           - CTG GCG GGG GAG GGC GAG GCC TGC CCG CTG CT - #G GGC AGC CCG GGC GCT         2225                                                                          Leu Ala Gly Glu Gly Glu Ala Cys Pro Leu Le - #u Gly Ser Pro Gly Ala           #               710                                                           - GGG CGA AAT AGC GTC CTC TTC CTC CCC GTG GA - #C CCC GAG GAC TCG CCC         2273                                                                          Gly Arg Asn Ser Val Leu Phe Leu Pro Val As - #p Pro Glu Asp Ser Pro           #           725                                                               - CTT GGC AGC AGC ACC CCC ATG GCG TCT CCT GA - #C CTC CTT CCA GAG GAC         2321                                                                          Leu Gly Ser Ser Thr Pro Met Ala Ser Pro As - #p Leu Leu Pro Glu Asp           #       740                                                                   - GTG AGG GAG CAC CTC GAA GGC TTG ATG CTC TC - #G CTC TTC GAG CAG AGT         2369                                                                          Val Arg Glu His Leu Glu Gly Leu Met Leu Se - #r Leu Phe Glu Gln Ser           #   755                                                                       - CTG AGC TGC CAG GCC CAG GGG GGC TGC AGT AG - #A CCC GCC ATG GTC CTC         2417                                                                          Leu Ser Cys Gln Ala Gln Gly Gly Cys Ser Ar - #g Pro Ala Met Val Leu           760                 7 - #65                 7 - #70                 7 -       #75                                                                           - ACA GAC CCA CAC ACG CCC TAC GAG GAG GAG CA - #G CGG CAG TCA GTG CAG         2465                                                                          Thr Asp Pro His Thr Pro Tyr Glu Glu Glu Gl - #n Arg Gln Ser Val Gln           #               790                                                           - TCT GAC CAG GGC TAC ATC TCC AGG AGC TCC CC - #G CAG CCC CCC GAG GGA         2513                                                                          Ser Asp Gln Gly Tyr Ile Ser Arg Ser Ser Pr - #o Gln Pro Pro Glu Gly           #           805                                                               - CTC ACG GAA ATG GAG GAA GAG GAG GAA GAG GA - #G CAG GAC CCA GGG AAG         2561                                                                          Leu Thr Glu Met Glu Glu Glu Glu Glu Glu Gl - #u Gln Asp Pro Gly Lys           #       820                                                                   - CCG GCC CTG CCA CTC TCT CCC GAG GAC CTG GA - #G AGC CTG AGG AGC CTC         2609                                                                          Pro Ala Leu Pro Leu Ser Pro Glu Asp Leu Gl - #u Ser Leu Arg Ser Leu           #   835                                                                       - CAG CGG CAG CTG CTT TTC CGC CAG CTG CAG AA - #G AAC TCG GGC TGG GAC         2657                                                                          Gln Arg Gln Leu Leu Phe Arg Gln Leu Gln Ly - #s Asn Ser Gly Trp Asp           840                 8 - #45                 8 - #50                 8 -       #55                                                                           - ACG ATG GGG TCA GAG TCA GAG GGG CCC AGT GC - #A TGA GGGCGGCTCC              2703                                                                          Thr Met Gly Ser Glu Ser Glu Gly Pro Ser Al - #a  *                            #               865                                                           - CCAGGGACCG CCCAGATCCC AGCTTTGAGA GAGGAGTGTG TGTGCACGTA TT - #CATCTGTG       2763                                                                          - TGTACATGTC TGCATGTGTA TATGTTCGTG TGTGAAATGT AGGCTTTAAA AT - #GTAAATGT       2823                                                                          - CTGGATTTTA ATCCCAGGCA TCCCTCCTAA CTTTTCTTTG TGCAGCGGTC TG - #GTTATCGT       2883                                                                          - CTATCCCCAG GGGAATCCAC ACAGCCCGCT CCCAGGAGCT AATGGTAGAG CG - #TCCTTGAG       2943                                                                          - GCTCCATTAT TCGTTCATTC AGCATTTATT GTGCACCTAC TATGTGGCGG GC - #ATTTGGGA       3003                                                                          - TACCAAGATA AATTGCATGC GGCATGGCCC CAGCCATGAA GGAACTTAAC CG - #CTAGTGCC       3063                                                                          - GAGGACACGT TAAACGAACA GGATGGGCCG GGCACGGTGG CTCACGCCTG TA - #ATCCCAGC       3123                                                                          - ACACTGGGAG GCCGAGGCAG GTGGATCACT CTGAGGTCAG GAGTTTGAGC CA - #GCCTGGCC       3183                                                                          #  3223            GGAA TTCGAGCTCG GTACCCGGGG                                 - (2) INFORMATION FOR SEQ ID NO:10:                                           -      (i) SEQUENCE CHARACTERISTICS:                                                    (A) LENGTH:  866 ami - #no acids                                              (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                - Met Gly Ala Ala Arg Ser Pro Pro Ser Ala Va - #l Pro Gly Pro Leu Leu         #                 15                                                          - Gly Leu Leu Leu Leu Leu Leu Gly Val Leu Al - #a Pro Gly Gly Ala Ser         #             30                                                              - Leu Arg Leu Leu Asp His Arg Ala Leu Val Cy - #s Ser Gln Pro Gly Leu         #         45                                                                  - Asn Cys Thr Val Lys Asn Ser Thr Cys Leu As - #p Asp Ser Trp Ile His         #     60                                                                      - Pro Arg Asn Leu Thr Pro Ser Ser Pro Lys As - #p Leu Gln Ile Gln Leu         # 80                                                                          - His Phe Ala His Thr Gln Gln Gly Asp Leu Ph - #e Pro Val Ala His Ile         #                 95                                                          - Glu Trp Thr Leu Gln Thr Asp Ala Ser Ile Le - #u Tyr Leu Glu Gly Ala         #           110                                                               - Glu Leu Ser Val Leu Gln Leu Asn Thr Asn Gl - #u Arg Leu Cys Val Arg         #       125                                                                   - Phe Glu Phe Leu Ser Lys Leu Arg His His Hi - #s Arg Arg Trp Arg Phe         #   140                                                                       - Thr Phe Ser His Phe Val Val Asp Pro Asp Gl - #n Glu Tyr Glu Val Thr         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Val His His Leu Pro Lys Pro Ile Pro Asp Gl - #y Asp Pro Asn His Gln         #               175                                                           - Ser Lys Asn Phe Leu Val Pro Asp Cys Glu Hi - #s Ala Arg Met Lys Val         #           190                                                               - Thr Thr Pro Cys Met Ser Ser Gly Ser Leu Tr - #p Asp Pro Asn Ile Thr         #       205                                                                   - Val Glu Thr Leu Glu Ala His Gln Leu Arg Va - #l Ser Phe Thr Leu Trp         #   220                                                                       - Asn Glu Ser Thr His Tyr Gln Ile Leu Leu Th - #r Ser Phe Pro His Met         225                 2 - #30                 2 - #35                 2 -       #40                                                                           - Glu Asn His Ser Cys Phe Glu His Met His Hi - #s Ile Pro Ala Pro Arg         #               255                                                           - Pro Glu Glu Phe His Gln Arg Ser Asn Val Th - #r Leu Thr Leu Arg Asn         #           270                                                               - Leu Lys Gly Cys Cys Arg His Gln Val Gln Il - #e Gln Pro Phe Phe Ser         #       285                                                                   - Ser Cys Leu Asn Asp Cys Leu Arg His Ser Al - #a Thr Val Ser Cys Pro         #   300                                                                       - Glu Met Pro Asp Thr Pro Glu Pro Ile Pro As - #p Tyr Met Pro Leu Trp         305                 3 - #10                 3 - #15                 3 -       #20                                                                           - Val Tyr Trp Phe Ile Thr Gly Ile Ser Ile Le - #u Leu Val Gly Ser Val         #               335                                                           - Ile Leu Leu Ile Val Cys Met Thr Trp Arg Le - #u Ala Gly Pro Gly Ser         #           350                                                               - Glu Lys Tyr Ser Asp Asp Thr Lys Tyr Thr As - #p Gly Leu Pro Ala Ala         #       365                                                                   - Asp Leu Ile Pro Pro Pro Leu Lys Pro Arg Ly - #s Val Trp Ile Ile Tyr         #   380                                                                       - Ser Ala Asp His Pro Leu Tyr Val Asp Val Va - #l Leu Lys Phe Ala Gln         385                 3 - #90                 3 - #95                 4 -       #00                                                                           - Phe Leu Leu Thr Ala Cys Gly Thr Glu Val Al - #a Leu Asp Leu Leu Glu         #               415                                                           - Glu Gln Ala Ile Ser Glu Ala Gly Val Met Th - #r Trp Val Gly Arg Gln         #           430                                                               - Lys Gln Glu Met Val Glu Ser Asn Ser Lys Il - #e Ile Val Leu Cys Ser         #       445                                                                   - Arg Gly Thr Arg Ala Lys Trp Gln Ala Leu Le - #u Gly Arg Gly Ala Pro         #   460                                                                       - Val Arg Leu Arg Cys Asp His Gly Lys Pro Va - #l Gly Asp Leu Phe Thr         465                 4 - #70                 4 - #75                 4 -       #80                                                                           - Ala Ala Met Asn Met Ile Leu Pro Asp Phe Ly - #s Arg Pro Ala Cys Phe         #               495                                                           - Gly Thr Tyr Val Val Cys Tyr Phe Ser Glu Va - #l Ser Cys Asp Gly Asp         #           510                                                               - Val Pro Asp Leu Phe Gly Ala Ala Pro Arg Ty - #r Pro Leu Met Asp Arg         #       525                                                                   - Phe Glu Glu Val Tyr Phe Arg Ile Gln Asp Le - #u Glu Met Phe Gln Pro         #   540                                                                       - Gly Arg Met His Arg Val Gly Glu Leu Ser Gl - #y Asp Asn Tyr Leu Arg         545                 5 - #50                 5 - #55                 5 -       #60                                                                           - Ser Pro Gly Gly Arg Gln Leu Arg Ala Ala Le - #u Asp Arg Phe Arg Asp         #               575                                                           - Trp Gln Val Arg Cys Pro Asp Trp Phe Glu Cy - #s Glu Asn Leu Tyr Ser         #           590                                                               - Ala Asp Asp Gln Asp Ala Pro Ser Leu Asp Gl - #u Glu Val Phe Glu Glu         #       605                                                                   - Pro Leu Leu Pro Pro Gly Thr Gly Ile Val Ly - #s Arg Ala Pro Leu Val         #   620                                                                       - Arg Glu Pro Gly Ser Gln Ala Cys Leu Ala Il - #e Asp Pro Leu Val Gly         625                 6 - #30                 6 - #35                 6 -       #40                                                                           - Glu Glu Gly Gly Ala Ala Val Ala Lys Leu Gl - #u Pro His Leu Gln Pro         #               655                                                           - Arg Gly Gln Pro Ala Pro Gln Pro Leu His Th - #r Leu Val Leu Ala Ala         #           670                                                               - Glu Glu Gly Ala Leu Val Ala Ala Val Glu Pr - #o Gly Pro Leu Ala Asp         #       685                                                                   - Gly Ala Ala Val Arg Leu Ala Leu Ala Gly Gl - #u Gly Glu Ala Cys Pro         #   700                                                                       - Leu Leu Gly Ser Pro Gly Ala Gly Arg Asn Se - #r Val Leu Phe Leu Pro         705                 7 - #10                 7 - #15                 7 -       #20                                                                           - Val Asp Pro Glu Asp Ser Pro Leu Gly Ser Se - #r Thr Pro Met Ala Ser         #               735                                                           - Pro Asp Leu Leu Pro Glu Asp Val Arg Glu Hi - #s Leu Glu Gly Leu Met         #           750                                                               - Leu Ser Leu Phe Glu Gln Ser Leu Ser Cys Gl - #n Ala Gln Gly Gly Cys         #       765                                                                   - Ser Arg Pro Ala Met Val Leu Thr Asp Pro Hi - #s Thr Pro Tyr Glu Glu         #   780                                                                       - Glu Gln Arg Gln Ser Val Gln Ser Asp Gln Gl - #y Tyr Ile Ser Arg Ser         785                 7 - #90                 7 - #95                 8 -       #00                                                                           - Ser Pro Gln Pro Pro Glu Gly Leu Thr Glu Me - #t Glu Glu Glu Glu Glu         #               815                                                           - Glu Glu Gln Asp Pro Gly Lys Pro Ala Leu Pr - #o Leu Ser Pro Glu Asp         #           830                                                               - Leu Glu Ser Leu Arg Ser Leu Gln Arg Gln Le - #u Leu Phe Arg Gln Leu         #       845                                                                   - Gln Lys Asn Ser Gly Trp Asp Thr Met Gly Se - #r Glu Ser Glu Gly Pro         #   860                                                                       - Ser Ala                                                                     865                                                                           __________________________________________________________________________

What is claimed is:
 1. A method for suppressing rejection of a graftedsyngeneic or allogeneic organ or grafted tissue in a graft recipient,comprising transfecting the organ or tissue to be transplanted with aDNA encoding a soluble IL-17R selected from the group consisting of:(a)a protein comprising amino acids 1 through 322 of SEQ ID NO.: 2; (b) aprotein comprising amino acids 1 through 320 of SEQ ID NO.: 10; (c)proteins encoded by DNA molecules which hybridize to DNA's encoding theproteins of (a) or (b) under moderately stringent conditions, and whichbind IL-17; and (d) fragments of the proteins of (a), (b) or (c) whichencode proteins comprising an extracellular domain of the proteins ofSEQ ID NO.:2 or SEQ ID NO.:10, that bind IL-17,and engrafting the organor tissue in the recipient, wherein the expression of IL-17R by theengrafted organ or tissue results in suppression of rejection.
 2. Themethod according to claim 1, further comprising administering,simultaneously, separately or sequentially, a composition comprising anIntereukin-17 receptor (IL-17R) protein selected from the groupconsisting of:(a) a protein comprising amino acids 1 through 322 of SEQID NO.: 2; (b) a protein comprising amino acids 1 through 320 of SEQ IDNO.: 10; (c) proteins encoded by DNA molecules which hybndize to DNA'sencoding the proteins of (a) or (b) under moderately stringentconditions, and which bind IL-17; and (d) fragments of the proteinsselected from the group consisting of a protein comprising amino acids 1through 322 of SEQ ID NO.: 2; a protein comprising amino acids 1 through320 of SEQ ID NO.: 10; and proteins encoded by DNA molecules whichhybridize to DNAs encoding the proteins comprising amino acids 1 through322 of SEQ ID NO:2 or amino acids 1 through 320 of SEQ ID NO:10 undermoderately stringent conditions, which encode proteins comprising anextracellular domain of the proteins of SEQ ID NO.:2 or SEQ ID NO.:10,that bind IL-17,and a suitable diluent or carrier, to the recipient,wherein the expression of IL-17R by the engrafted organ or tissueresults in suppression of rejection.
 3. A method for suppressingrejection of a grafted syngeneic or allogeneic organ or grafted tissuein a graft recipient, comprising transfecting the organ or tissue to betransplanted with a DNA encoding a soluble IL-17R protein selected fromthe group consisting of:(a) a protein comprising amino acids 1 through322 of SEQ ID NO.: 2; (b) a protein comprising amino acids 1 through 320of SEQ ID NO.: 10; (c) a protein that is at least about 70% identical tothe proteins of (a) or (b), and that binds IL-17; and (d) fragments ofthe proteins of (a), (b) or (c) which encode proteins comprising anextracellular domain of the proteins of SEQ ID NO.:2 or SEQ ID NO.:10,that bind IL-17,and a suitable diluent or carrier,wherein said DNA istransferred ex vivo or through direct injection into the organ and theexpression of IL-17R by the engrafied organ or tissue results insuppression of rejection.
 4. The method according to claim 3, furthercomprising administering, simultaneously, separately or sequentially, acomposition comprising an Interleukin-17 receptor (IL-17R) proteinselected from the group consisting of:(a) a protein comprising aminoacids 1 through 322 of SEQ ID NO.: 2; (b) a protein comprising aminoacids 1 through 320 of SEQ ID NO.: 10; (c) proteins encoded by DNAmolecules which hybridize to DNA's encoding the proteins of (a) or (b)under moderately stringent conditions, and which bind IL-17; and (d)fragments of the proteins selected from the croup consisting of aprotein comprising amino acids 1 through 322 of SEQ ID NO.: 2; a proteincomprising amino acids 1 through 320 of SEQ ID NO.: 10; and proteinsencoded by DNA molecules which hybridize to DNAs encoding the proteinscomprising amino acids 1 through 322 of SEQ ID NO:2 or amino acids 1through 320 of SEQ ID NO:10 under moderately stringent conditions, whichencode proteins comprising an extracellular domain of the proteins ofSEQ ID NO.:2 or SEQ ID NO.:10, that bind IL-17,and a suitable diluent orcarrier, to the recipient, wherein the expression of IL-17R by theengrafted organ or tissue results in suppression of rejection.
 5. Themethod according to claim 3, further comprising administering,simultaneously, separately or sequentially, a composition comprising anInterleukin-17 receptor (IL-17R) protein selected from the groupconsisting of:(a) a protein comprising amino acids 1 through 322 of SEQID NO.: 2; (b) a protein comprising amino acids 1 through 320 of SEQ IDNO.: 10; (c) a protein that is at least about 70% identical to theproteins of (a) or (b), and that binds IL-17; and (d) fragments of theproteins selected from the group consisting of a protein comprisingamino acids 1 through 322 of SEQ ID NO.: 2; a protein comprising aminoacids 1 through 320 of SEQ ID NO.: 10; and a protein that is at leastabout 70% identical to the proteins comprnsing amino acids 1 through 322of SEQ ID NO:2 or amino acids 1 through 320 of SEQ ID NO:10, whichencode proteins comprising an extracellular domain of the proteins ofSEQ ID NO.:2 or SEQ ID NO.:10, that bind IL-17,and a suitable diluent orcarrier, to the recipient, wherein the expression of IL-17R by theengrafted organ or tissue results in suppression of rejection.
 6. Themethod according to claim 3, further comprising administering,simultaneously, separately or sequentially, a composition comprising anInterleukin-17 receptor (IL-17R) protein selected from the groupconsisting of:(a) a protein comprising amino acids 1 through 322 of SEQID NO: 2; and (b) a protein comprising amino acids 1 through 320 of SEQID NO: 10, and a suitable diluent or carrier, to the recipient,whereinthe expression of IL-17R by the engrafted organ or tissue results insuppression of rejection.
 7. A method for suppressing rejection of agrafted syngeneic or allogeneic organ or grafted tissue in a graftrecipient, comprising transfecting the organ or tissue to betransplanted with a DNA encoding a soluble IL-17R protein, selected fromthe group consisting of:(a) a protein comprising amino acids 1 through322 of SEQ ID NO.: 2; and (b) a protein comprising amino acids 1 through320 of SEQ ID NO.: 10, and a suitable diluent or carrier,wherein saidDNA is transferred ex vivo or through direct injection into the organand the expression of IL-17R by the engrafted organ or tissue results insuppression of rejection.
 8. The method according to claim 7, furthercomprising administering, simultaneously, separately or sequentially, acomposition comprising an Interleukin-17 receptor (IL-17R) proteinselected from the group consisting of:(a) a protein comprising aminoacids 1 through 322 of SEQ ID NO.: 2; (b) a protein comprising aminoacids 1 through 320 of SEQ ID NO.: 10; (c) proteins encoded by DNAmolecules which hybridize to DNA's encoding the proteins of (a) or (b)under moderately stringent conditions, and which bind IL-17; and (d)fragments of the proteins selected from the group consisting of aprotein comprising amino acids 1 through 322 of SEQ ID NO.: 2; a proteincomprising amino acids 1 through 320 of SEQ ID NO.: 10; and proteinsencoded by DNA molecules which hybridize to DNAs encoding the proteinscomprising amino acids 1 through 322 of SEQ ID NO:2 or amino acids 1through 320 of SEQ ID NO:10 under moderately stringent conditions, whichencode proteins comprising an extracellular domain of the proteins ofSEQ ID NO.:2 or SEQ ID NO.:10, that bind IL-17,and a suitable diluent orcarrier, to the recipient herein the expression of IL-17R by theengrafted organ or tissue results in sunpression of rejection.
 9. Themethod according to claim 7, further comprising administering,simultaneously, separately or sequentially, a composition comprising anInterleukin-17 receptor (IL-17R) protein selected from the groupconsisting of:(a) a protein comprising amino acids 1 through 322 of SEQID NO.: 2; (b) a protein comprising amino acids 1 through 320 of SEQ IDNO.: 10; (c) a protein that is at least about 70% identical to theproteins of (a) or (b), and that binds IL-17; and (d) fragments of theproteins selected from the group consisting of a protein comprisingamino acids 1 through 322 of SEQ ID NO.: 2; a protein comprising aminoacids 1 through 320 of SEQ ID NO.: 10; and a protein that is at leastabout 70% identical to the proteins comprising amino acids 1 through 322of SEQ ID NO:2 or amino acids 1 through 320 of SEQ ID NO:10, whichencode proteins comprising an extracellular domain of the proteins ofSEQ ID NO.:2 or SEQ ID NO.: 10, that bind IL-17,and a suitable diluentor carrier, to the recipient, wherein the expression of IL-17R by theengrafted organ or tissue results in suppression of rejection.
 10. Themethod according to claim 7, further comprising administering,simultaneously, separately or sequentially, a composition comprising anInterleukin-17 receptor (IL-17R) protein selected from the groupconsisting of:(a) a protein comprising amino acids 1 through 322 of SEQID NO.: 2; and (b) a protein comprising amino acids 1 through 320 of SEQID NO.: 10;and a suitable diluent or carrier, to the recipient, whereinthe expression of IL-17R by the engrafted organ or tissue results insuppression of rejection.